Compounds for the treatment of neurological or mitochondrial diseases

ABSTRACT

Compounds and methods are provided for the treatment of neurological or mitochondrial diseases, including epilepsy. In some embodiments, the compounds are substituted 1,4-naphthoquinones.

This application is a divisional application of U.S. patent applicationSer. No. 17/264,714 filed Jan. 29, 2021, which is a national phaseapplication under 35 U.S.C. § 371 of International Application No.PCT/US2019/043868, filed Jul. 29, 2019 which claims the benefit of U.S.Provisional Patent Application No. 62/711,575, filed Jul. 29, 2018, theentirety of each of which are incorporated herein by reference.

This invention was made with government support under Grant No.NS097047-01, ES015555, RR024485, GM103542, and RR029882 awarded by theNational Institutes of Health. The government has certain rights in theinvention.

BACKGROUND OF THE INVENTION 1. Field of the Invention

The present invention relates generally to the field of chemistry andmedicine. More particularly, it concerns compounds that may be used,e.g., for the treatment of neuronal disorders such as epilepsy.

2. Description of Related Art

Epilepsy is a widespread, debilitating disease with treatments thatsuffer from poor adherence, adverse side effects, or even inefficacy inpatients with various forms of medication-resistance epilepsy. Epilepsyis affects approximately 1-2% of the world's population and ischaracterized by the periodic and unpredictable occurrence of seizures(Bialer and White 2010).

The more than 25 anti-epileptic drugs (AEDs) on the market sufferseveral drawbacks, including side effects and adverse drug interactionsthat can lead to decreased patient quality of life and poor treatmentadherence (Franco et al. 2016; Loscher et al.). For example, commonlyprescribed valproic acid (VPA), a broad-spectrum AED used to treat allforms of seizures (Perucca E. 2002), is generally well tolerated butrequires high therapeutic doses and is associated with several sideeffects: acute hepatic failure, pancreatitis and teratogenesis (Lheureuxand Hantson, 2009). Thus, VPA is contra-indicated for young children(Stewart et al. 2010) and pregnant women (Alsdorf and Wyszynski, 2005),and can induce rapid decline in health in mitochondrial disease patients(Finsterer and Segall, 2010). Some more recently generated AEDs displaylowered toxicity and fewer adverse drug interactions but are onlymarginally (if at all) more efficacious than older drugs (Loscher andSchmidt, 2011).

Medication-resistant epilepsy presents a particularly difficult clinicalproblem. Indeed, ˜40% of patients experience medication-resistantepilepsy, which no currently marketed drug has been shown to control(Loscher and Schmidt, 2011; Mohanraj and Brodie, 2005). These patientsinclude up to 90% of pediatric patients with medication-resistantepilepsies due to genetic mitochondrial disorders, such as mitochondrialDNA depletion syndrome (MDS) (Finsterer and Scorza, 2017).

A poor understanding of how to target the underlying causes of epilepsyimpedes therapeutic breakthroughs (Bindoff and Engelsen, 2011; Bindoffand Engelsen, 2012; Loscher et al., 2013; Noebels et al. 2012).Additionally, previously identified compounds that have displayedencouraging results in vitro have often displayed poor pharmacokineticprofiles, limiting their potential for use as an anti-epilepsy drug(AED) (Rahn et al., 2013). Despite the fact that mitochondrialdysfunction is a major underlying cause of epilepsy, there are currentlyno AEDs for clinical use that target mitochondrial health andenergetics. Clearly, there is a need for new compounds for the treatmentof epilepsy and medication resistant epilepsy.

SUMMARY OF THE INVENTION

The present invention, in certain aspects, overcomes limitations in theprior art by providing compounds that may be used for the treatment ofneurological or metabolic diseases such as, e.g., epilepsy ormedication-resistant epilepsy. In some embodiments, and as shown in invivo experiments in the below examples, compounds are provided hereinthat display improved anti-seizure activity and improved pharmacokineticproperties. In some embodiments, compounds are provided that resulted inimproved efficacy and drug-like properties; for example, inclusion of a2-pentynylamine substituent or an isoamyloxy group on an1,4-naphthoquinone scaffold resulted in significant improvements inanti-seizure activity in zebrafish and improved pharmacokineticproperties in mice.

In some aspects, the present disclosure provides methods of treating aneurological disease or a mitochondrial disease in a mammalian subjectcomprising administering to a subject a therapeutically effective doseof a pharmaceutical composition comprising a compound of the formula:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of any of the last five groups;    -   Y is —NH— or —O—;    -   wherein when Y is —NH— and R₂ is hydrogen,        -   R₁ is C₁₋₈ alkyl, C₆₋₁₈ alkenyl, or C₆₋₁₂ aryl;            —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2, 3, or 4 and R_(a) is            C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂            heteroaryl, or a substituted version of any of these groups;        -   a group of the formula:

-   -   wherein:        -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,            substituted C₆₋₁₂ aryl, or —C(O)R_(d);            -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆ alkoxy,                C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a substituted                version of any of the last three groups; or        -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,            C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted version of            any of the last five groups; and        -   y is 0, 1, or 2; or    -   a group of the formula:

-   -   wherein:        -   z is 1, 2, or 3        -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆            alkylsulfonylamino, or a substituted version of any of the            last three groups; or    -   a group of the formula:

-   -   wherein:        -   a is 1, 2, or 3        -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted version            of any of either groups;            wherein when Y is —NH— and R₂ is not hydrogen; then:    -   R₁ is C₁₋₁₂ alkyl, C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂ aryl,        C₁₋₁₂ heteroaryl, or a substituted version of any of these        groups, or —Y′—X₃—R_(g), wherein:        -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a substituted            version of either group;        -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—; and        -   R_(g) is C₁₋₆ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂ aryl,            C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a substituted version of            any of these groups; or    -   wherein when Y is —O—,        -   R₁ is C₁₋₁₈ alkyl, C₁₋₁₈ alkenyl, C₁₋₁₈ alkynyl, C₇₋₁₈            aralkyl, or a substituted version of any of these groups;        -   or            -   a group of the formula:

-   -   -   -   wherein:                -   R_(b) is hydrogen, C₁₋₈ alkyl, substituted C₁₋₈                    alkyl, C₆₋₁₂ aryl, substituted C₆₋₁₂ aryl, or                    —C(O)R_(d);                -    wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or                -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                    alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a                    substituted version of any of the last five groups;                    and                -   y is 0, 1, 2, or 3;                    or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is further defined as:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of any of the last five groups;    -   Y is —NH— or —O—;    -   wherein when Y is —NH— and R₂ is hydrogen,        -   R₁ is C₆₋₁₈ alkenyl; —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2,            3, or 4 and R_(a) is C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,            C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted version of            any of these groups;        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,                substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, or 2; or        -   a group of the formula:

-   -   -   wherein:            -   z is 1, 2, or 3            -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆                alkylsulfonylamino, or a substituted version of any of                the last three groups; or        -   a group of the formula:

-   -   -   wherein:            -   a is 1, 2, or 3            -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted                version of any of either groups;

    -   wherein when Y is —NH— and R₂ is not hydrogen; then:        -   R₁ is C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂            heteroaryl, or a substituted version of any of these groups,            or —Y′—X₃—R_(g), wherein:            -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a substituted                version of either group;            -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—; and            -   R_(g) is C₁₋₆ alkyl C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a substituted                version of any of these groups; or

    -   wherein when Y is —O—,        -   R₁ is C₁₋₁₈ alkyl, substituted C₁₋₁₈ alkyl, C₁₋₁₈ alkenyl,            substituted C₁₋₁₈ alkenyl, C₁₋₁₈ alkynyl, or substituted            C₁₋₁₈ alkynyl; or        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,                substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, 2, or 3;                or a pharmaceutically acceptable salt thereof.

In some embodiments, Y is —NH—, R₂ is hydrogen, and R₁ is C₁₋₈ alkyl,such isopentyl.

In some embodiments, the disease is a neurological disease such asepilepsy, bipolar disorder or the manic phase of bipolar disorder,headaches, migraines, a traumatic brain injury, Parkinson's disease,Alzheimer's disease, Huntington's disease, Friedereich's Ataxia, oroptic atrophy. In some embodiments, the neurological disease is epilepsysuch as medication-resistant epilepsy. In other embodiments, the diseaseis a mitochondrial disease such as mitochondrial DNA depletion syndromeor dysfunctional mitochondrial respiratory chain disorder.

In some embodiments, the pharmaceutical composition is formulated fororal, sublingual, intranasal, intravenous, subcutaneous, parenteral,inhalation, or aerosol delivery; thus, in some embodiments thepharmaceutical composition may be administered orally, sublingually,intranasally, intravenously, subcutaneously, parenterally, or viainhalation or aerosol. In some embodiments, the subject is a human. Insome embodiments, the compound has the structure:

wherein:

-   -   R₂, R₃, X₁ and X₂ are as defined above; and    -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,        substituted C₆₋₁₂ aryl, or —C(O)R_(d);        -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆ alkoxy, C₁₋₆            alkylamino, or C₁₋₈ dialkylamino, or a substituted version            of any of the last three groups; or    -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂        aryl, C₁₋₁₂ heteroaryl, or a substituted version of any of the        last five groups; and    -   y is 0, 1, or 2;        or a pharmaceutically acceptable salt thereof.

In some embodiments, X₁ and X₂ are ═O. In some embodiments, R₃ ishydrogen. In some embodiments, R₄ is hydrogen. In some embodiments, R₁is —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2, 3, or 4 and R_(a) is C₁₋₈alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or asubstituted version of any of these groups. In some embodiments, R₁ is—(CH₂)_(y1)—C≡C—C₁₋₈ alkyl, wherein y1=1-2. In some embodiments, R₁ is—(CH₂)_(y1)—C≡C—C₁₋₃ alkyl, wherein y1=1-2 such as—(CH₂)_(y1)—C≡C—(CH₂)_(y2)—CH₃, wherein y2=1-6. In some embodiments, R₁is

CH₃, —(CH₂)₂—C≡C—(CH₂)₂—CH₃, —CH₂—C≡C—(CH₂)₂—CH₃, —CH₂—C≡C—(CH₂)₃—CH₃,—CH₂—C≡C—(CH₂)₆—CH₃, or —CH₂—C≡C—CH₂—CH═CH—CH₃. In some embodiments,R_(f) is substituted C₆₋₁₂ aryl or C₁₋₁₂ heteroaryl. In someembodiments, R_(f) is substituted with a —F, —OCH₃, —CF₃, or —NHS(O)₂CH₃group.

In some embodiments, the compound is further defined as:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is:

or a pharmaceutically acceptable salt thereof.

In other embodiments, the compound is:

In some embodiments, R₁ is

In some embodiments, R₁ is

In some embodiments, the compound is:

or a pharmaceutically acceptable salt thereof. In some embodiments, thecompound is formulated with a pharmaceutically acceptable excipient.

In another aspect, the present disclosure provides pharmaceuticalcompositions comprising:

-   -   (A) a compound of the formula:

-   -   wherein:        -   X₁ and X₂ are each independently oxo or hydroxy;        -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl,            C₁₋₆ alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,            —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted            version thereof;        -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,            aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆            alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted            version of any of the last five groups;        -   Y is —NH— or —O—;        -   wherein when Y is —NH— and R₂ is hydrogen,            -   R₁ is C₁₋₈ alkyl, C₆₋₁₈ alkenyl, or C₆₋₁₂ aryl;                —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2, 3, or 4 and                R_(a) is C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, or a substituted version of any                of these groups;            -   a group of the formula:

-   -   -   -   wherein:                -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂                    aryl, substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -    wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or                -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                    alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a                    substituted version of any of the last five groups;                    and                -   y is 0, 1, or 2; or            -   a group of the formula:

-   -   -   -   wherein:                -   z is 1, 2, or 3                -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆                    alkylsulfonylamino, or a substituted version of any                    of the last three groups; or            -   a group of the formula:

-   -   -   -   wherein:                -   a is 1, 2, or 3                -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted                    version of any of either groups;

        -   wherein when Y is —NH— and R₂ is not hydrogen; then:            -   R₁ is C₁₋₁₂ alkyl, C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, or a substituted version of any                of these groups, or —Y′—X₃—R_(g), wherein:                -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a                    substituted version of either group;                -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—;                    and                -   R_(g) is C₁₋₆ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,                    C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a                    substituted version of any of these groups; or

        -   wherein when Y is —O—,            -   R₁ is C₁₋₁₈ alkyl, C₁₋₁₈ alkenyl, C₁₋₁₈ alkynyl, C₇₋₁₈                aralkyl, or a substituted version of any of these                groups;            -   or                -   a group of the formula:

-   -   -   -   -   wherein:                -    R_(b) is hydrogen, C₁₋₈ alkyl, substituted C₁₋₈                    alkyl, C₆₋₁₂ aryl, substituted C₆₋₁₂ aryl, or                    —C(O)R_(d);                -    wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or                -    R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                    alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a                    substituted version of any of the last five groups;                    and                -    y is 0, 1, 2, or 3;

    -   or a pharmaceutically acceptable salt thereof; and

    -   (B) an excipient.

In some embodiments, the compound is further defined as:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of the last five groups;    -   Y is —NH— or —O—;    -   wherein when Y is —NH— and R₂ is hydrogen,        -   R₁ is C₆₋₁₈ alkenyl; —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2,            3, or 4 and R_(a) is C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,            C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted version of            any of these groups;        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,                substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, or 2; or        -   a group of the formula:

-   -   -   wherein:            -   z is 1, 2, or 3            -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆                alkylsulfonylamino, or a substituted version of any of                the last three groups; or        -   a group of the formula:

-   -   -   wherein:            -   a is 1, 2, or 3            -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted                version of any of either groups;

    -   wherein when Y is —NH— and R₂ is not hydrogen; then:        -   R₁ is C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂            heteroaryl, or a substituted version of any of these groups,            or —Y′—X₃—R_(g), wherein:            -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a substituted                version of either group;            -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—; and            -   R_(g) is C₁₋₆ alkyl C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a substituted                version of either group; or

    -   wherein when Y is —O—,        -   R₁ is C₁₋₁₈ alkynyl or substituted C₁₋₁₈ alkynyl;        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is hydrogen, C₁₋₈ alkyl, substituted C₁₋₈ alkyl,                C₆₋₁₂ aryl, substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, 2, or 3; or                or a pharmaceutically acceptable salt thereof.

In some embodiments, the pharmaceutical composition is administeredorally, sublingually, intranasally, intravenously, subcutaneously,parenterally, via inhalation, or aerosol. In some embodiments, thecompound is the compound described herein. In some embodiments, thepharmaceutical composition is formulated as a unit dose.

In some aspects the present disclosure provides pharmaceuticalcompositions comprising:

-   -   (A) a compound of the formula:

-   -   wherein:        -   X₁ and X₂ are each independently oxo or hydroxy;        -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl,            C₁₋₆ alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,            —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted            version thereof;        -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,            aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆            alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted            version of any of the last five groups;        -   Y is —NH— or —O—;        -   wherein when Y is —NH— and R₂ is hydrogen,            -   R₁ is C₁₋₈ alkyl, C₆₋₁₈ alkenyl, or C₆₋₁₂ aryl;                —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2, 3, or 4 and                R_(a) is C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, or a substituted version of any                of these groups;            -   a group of the formula:

-   -   -   -   wherein:                -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂                    aryl, substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -    wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or                -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                    alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a                    substituted version of any of the last five groups;                    and                -   y is 0, 1, or 2; or            -   a group of the formula:

-   -   -   -   wherein:                -   z is 1, 2, or 3                -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆                    alkylsulfonylamino, or a substituted version of any                    of the last three groups; or            -   a group of the formula:

-   -   -   -   wherein:                -   a is 1, 2, or 3                -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted                    version of any of either groups;

        -   wherein when Y is —NH— and R₂ is not hydrogen; then:            -   R₁ is C₁₋₁₂ alkyl, C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, or a substituted version of any                of these groups, or —Y′—X₃—R_(g), wherein:                -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a                    substituted version of either group;                -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—;                    and                -   R_(g) is C₁₋₆ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,                    C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a                    substituted version of any of these groups; or

        -   wherein when Y is —O—,            -   R₁ is C₁₋₁₈ alkyl, C₁₋₁₈ alkenyl, C₁₋₁₈ alkynyl, C₇₋₁₈                aralkyl, or a substituted version of any of these                groups;            -   or                -   a group of the formula:

-   -   -   -   -   wherein:                -    R_(b) is hydrogen, C₁₋₈ alkyl, substituted C₁₋₈                    alkyl, C₆₋₁₂ aryl, substituted C₆₋₁₂ aryl, or                    —C(O)R_(d);                -    wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or                -    R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                    alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a                    substituted version of any of the last five groups;                    and                -    y is 0, 1, 2, or 3;

    -   or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is further defined as:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of any of the last five groups;    -   Y is —NH— or —O—;    -   wherein when Y is —NH— and R₂ is hydrogen,        -   R₁ is C₆₋₁₈ alkenyl; —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2,            3, or 4 and R_(a) is C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,            C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted version of            any of these groups;        -   a group of the formula:

-   -   -   -   wherein:                -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂                    aryl, substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -    wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or                -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                    alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a                    substituted version of any of the last five groups;                    and                -   y is 0, 1, or 2; or            -   a group of the formula:

-   -   -   wherein:            -   z is 1, 2, or 3            -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆                alkylsulfonylamino, or a substituted version of any of                the last three groups; or        -   a group of the formula:

-   -   -   wherein:            -   a is 1, 2, or 3            -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted                version of any of either groups;        -   wherein when Y is —NH— and R₂ is not hydrogen; then:            -   R₁ is C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂                heteroaryl, or a substituted version of any of these                groups, or —Y′—X₃—R_(g), wherein:                -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a                    substituted version of either group;                -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—;                    and                -   R_(g) is C₁₋₆ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,                    C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a                    substituted version of any of these groups; or        -   wherein when Y is —O—,            -   R₁ is C₁₋₁₈ alkyl, substituted C₁₋₁₈ alkyl, C₁₋₁₈                alkenyl, substituted C₁₋₁₈ alkenyl, C₁₋₁₈ alkynyl, and                substituted C₁₋₁₈ alkynyl; or                -   a group of the formula:

-   -   -   -   -   wherein:                -    R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂                    aryl, substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -    wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or                -    R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                    alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a                    substituted version of any of the last five groups;                    and                -    y is 0, 1, 2, or 3;                    or a pharmaceutically acceptable salt thereof.

In some embodiments, Y is —NH—, R₂ is hydrogen, and R₁ is C₁₋₈ alkyl,such as isopentyl.

In some embodiments, the compound has the structure:

wherein:

-   -   R₂, R₃, X₁ and X₂ are as defined above; and    -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,        substituted C₆₋₁₂ aryl, or —C(O)R_(d);        -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆ alkoxy, C₁₋₆            alkylamino, or C₁₋₈ dialkylamino, or a substituted version            of any of the last three groups; or    -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂        aryl, C₁₋₁₂ heteroaryl, or a substituted version of any of the        last five groups; and    -   y is 0, 1, or 2;        or a pharmaceutically acceptable salt thereof.

In some embodiments, X₁ and X₂ are ═O. In some embodiments, R₃ ishydrogen. In some embodiments, R₄ is hydrogen. In some embodiments, R₁is —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2, 3, or 4 and R_(a) is C₁₋₈alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or asubstituted version of any of these groups. In some embodiments, R₁ is—(CH₂)_(y1)—C≡C—C₁₋₈ alkyl, wherein y1=1-2. In some embodiments, R₁ is—(CH₂)_(y1)—C≡C—C₁₋₃ alkyl, wherein y1=1-2. In some embodiments, R₁ is—(CH₂)_(y1)—C≡C—(CH₂)_(y2)—CH₃, wherein y2=1-6. In some embodiments, R₁is

—(CH₂)₂—C≡C—CH₂—CH₃—CH₃, —(CH₂)₂—C≡C—(CH₂)₂—CH₃, —CH₂—C≡C—(CH₂)₂—CH₃,—CH₂—C≡C—(CH₂)₃—CH₃, —CH₂—C≡C—(CH₂)₆—CH₃, or —CH₂—C≡C—CH₂—CH═CH—CH₃. Insome embodiments, R_(f) is substituted C₆₋₁₂ aryl or C₁₋₁₂ heteroaryl.In some embodiments, R_(f) is substituted with a —F, —OCH₃, —CF₃, or—NHS(O)₂CH₃ group. In some embodiments, the compound is further definedas:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments, R₁ is

In some embodiments, the compound is:

or a pharmaceutically acceptable salt thereof.

In still yet another aspect, the present disclosure provides compoundshaving the structure:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of any of the last five groups;    -   Y is —NH— or —O—;    -   wherein when Y is —NH— and R₂ is hydrogen,        -   R₁ is isopentyl, C₆₋₁₈ alkenyl, or C₆₋₁₂ aryl;            —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2, 3, or 4 and R_(a) is            C₂₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₁₋₁₂ heteroaryl, or            a substituted version of any of these groups;        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,                substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, or 2; or        -   a group of the formula:

-   -   -   wherein:            -   z is 1, 2, or 3            -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆                alkylsulfonylamino, or a substituted version of any of                the last three groups; or        -   a group of the formula:

-   -   -   wherein:            -   a is 1, 2, or 3            -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted                version of any of either groups;

    -   wherein when Y is —NH— and R₂ is not hydrogen; then:        -   R₁ is C₁₋₁₂ alkyl, C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂ aryl,            C₁₋₁₂ heteroaryl, or a substituted version of any of these            groups, or —Y′—X₃—R_(g), wherein:            -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a substituted                version of either group;            -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—; and            -   R_(g) is C₁₋₆ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a substituted                version of any of these groups; or

    -   wherein when Y is —O—,        -   R₁ is C₁₋₁₈ alkyl, C₁₋₁₈ alkenyl, C₁₋₁₈ alkynyl, C₇₋₁₈            aralkyl, or a substituted version of any of these groups;        -   or            -   a group of the formula:

-   -   -   -   wherein:                -   R_(b) is hydrogen, C₁₋₈ alkyl, substituted C₁₋₈                    alkyl, C₆₋₁₂ aryl, substituted C₆₋₁₂ aryl, or                    —C(O)R_(d);                -    wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or                -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                    alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a                    substituted version of any of the last five groups;                    and                -   y is 0, 1, 2, or 3;                    or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is further defined as:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of the last five groups;    -   Y is —NH— or —O—;    -   wherein when Y is —NH— and R₂ is hydrogen,        -   R₁ is C₆₋₁₈ alkenyl, substituted C₆₋₁₈ alkenyl, or            —(CH₂)_(x)C≡CR_(a), wherein: x is 1, 2, 3, or 4 and R_(a) is            C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₁₋₁₂ heteroaryl, or a            substituted version of any of these groups;        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,                substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, or 2; or        -   a group of the formula:

-   -   -   wherein:            -   z is 1, 2, or 3            -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆                alkylsulfonylamino, or a substituted version of any of                the last three groups; or        -   a group of the formula:

-   -   -   wherein:            -   a is 1, 2, or 3            -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted                version of any of either groups;

    -   wherein when Y is —NH— and R₂ is not hydrogen; then:        -   R₁ is C₆₋₁₂ alkenyl, C₇₋₁₂ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂            heteroaryl, or a substituted version of any of these groups,            or —Y′—X₃—R_(g), wherein:            -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a substituted                version of either group;            -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—; and            -   R_(g) is C₁₋₆ alkyl C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a substituted                version of either group; or

    -   wherein when Y is —O—,        -   R₁ is C₁₋₁₈ alkynyl or substituted C₁₋₁₈ alkynyl; or        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,                substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, 2, or 3; or                or a pharmaceutically acceptable salt thereof.

In some embodiments, the compounds are further defined as:

wherein:

-   -   R₂, R₃, X₁ and X₂ are as defined above; and    -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,        substituted C₆₋₁₂ aryl, or —C(O)R_(d);        -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆ alkoxy, C₁₋₆            alkylamino, or C₁₋₈ dialkylamino, or a substituted version            of any of the last three groups; or    -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂        aryl, C₁₋₁₂ heteroaryl, or a substituted version of any of the        last five groups; and    -   y is 0, 1, or 2;        or a pharmaceutically acceptable salt thereof.

In other embodiments, the compounds are further defined as:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of the last five groups;    -   Y is —NH—;    -   R₁ is C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂        heteroaryl, or a substituted version of any of these groups, or        —Y′—X₃—R_(g), wherein:        -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a substituted            version of either group;        -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—; and        -   R_(g) is C₁₋₆ alkyl C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂ aryl,            C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a substituted version of            either group; or            or a pharmaceutically acceptable salt thereof.

In other embodiments, the compounds are further defined as:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of the last five groups;    -   Y is —O—;    -   R₁ is C₁₋₁₈ alkynyl and substituted C₁₋₁₈ alkynyl; or    -   a group of the formula:

-   -   wherein:        -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,            substituted C₆₋₁₂ aryl, or —C(O)R_(d);            -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆ alkoxy,                C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a substituted                version of any of the last three groups; or        -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,            C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted version of            any of the last five groups; and        -   y is 0, 1, 2, or 3; or            or a pharmaceutically acceptable salt thereof.

In some embodiments, X₁ and X₂ are ═O. In some embodiments, R₃ ishydrogen. In some embodiments, R₄ is hydrogen. In some embodiments, R₁is —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2, 3, or 4 and R_(a) is C₁₋₈alkenyl, C₁₋₈ alkynyl, C₁₋₁₂ heteroaryl, or a substituted version of anyof these groups. In some embodiments, R_(f) is substituted C₆₋₁₂ aryl orC₁₋₁₂ heteroaryl such as where R_(f) is substituted with a —F, —OCH₃,—CF₃, or —NHS(O)₂CH₃ group. In some embodiments, the compound is furtherdefined as:

-   -   or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is

In other embodiments, the compound is:

In some embodiments, R₁ is

In some embodiments, the compound is not2-((3-Phenylprop-2-yn-1-yl)amino)naphthalene-1,4-dione,2-(Isopentylamino)naphthalene-1,4-dione,2-(Isopentyloxy)naphthalene-1,4-dione, 2-Butoxynaphthalene-1,4-dione, or2-(Benzyloxy)naphthalene-1,4-dione.

In still yet another aspect, the present disclosure provides compoundsof the formula:

-   -   or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is

or a pharmaceutically acceptable salt thereof. In some embodiments, thecompound is comprised in a pharmaceutical composition or in apharmaceutically acceptable excipient.

As used herein the specification, “a” or “an” may mean one or more. Asused herein in the claim(s), when used in conjunction with the word“comprising”, the words “a” or “an” may mean one or more than one.

The use of the term “or” in the claims is used to mean “and/or” unlessexplicitly indicated to refer to alternatives only or the alternativesare mutually exclusive, although the disclosure supports a definitionthat refers to only alternatives and “and/or.” As used herein “another”may mean at least a second or more.

Throughout this application, the term “about” is used to indicate that avalue includes the inherent variation of error for the device, themethod being employed to determine the value, or the variation thatexists among the study subjects.

Other objects, features and advantages of the present disclosure willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and the specificexamples, while indicating specific embodiments of the invention, aregiven by way of illustration only, since various changes andmodifications within the spirit and scope of the invention will becomeapparent to those skilled in the art from this detailed description.Note that simply because a particular compound is ascribed to oneparticular generic formula doesn't mean that it cannot also belong toanother generic formula.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings form part of the present specification and areincluded to further demonstrate certain aspects of the presentinvention. The invention may be better understood by reference to one ormore of these drawings in combination with the detailed description ofspecific embodiments presented herein.

FIG. 1A-B: FIG. 1A, PK data for 1 in CD-1 mice. The drug wasadministered at 20 mg/kg via i.p. route in adult mice. Concentrationswere recorded using blood draws at six time points and are displayed inred, green, and blue for each individual animal. FIG. 1B, Plasmastability data for 1. In CD-1 mouse plasma ex vivo, the drug iscompletely stable at room temperature over the course of two hours. Dataexpressed as mean±SEM for three samples; differences at 0 hr and 2 hrare statistically insignificant.

FIG. 2 : Classes of compounds examined as potential AEDs.

FIG. 3 : Chemical syntheses of compounds examined as potential AEDs.

FIG. 4 : Dose-response curves for selected compounds, demonstratingtheir ability to protect HT22 cells from Glu-induced oxytocic celldeath. Cytoprotection data are expressed as percentage of viable Glu-and drug-treated cells as determined by relative resorufin fluorescencedata normalized to fluorescence measured for untreated cells (100%viability) and Glu-treated cells unexposed to drug compounds (0%viability). Curves were fitted from 8-10 point titrations (n=2-4) withdata points showing mean cytoprotection %±SEM.

FIGS. 5A-B: Compounds 3 (FIG. 5A) and 25 (FIG. 5B) decrease seizures ina zebrafish model of epilepsy in a dose-dependent manner, and havebetter efficacy than our previous lead, 1. Zebrafish at 7 dpf weretreated with 1.25-20 μM of 3 or 25 with pentylenetetrazole (PTZ, aconvulsant agent). Seizure activity is expressed as a fraction ofmonitored fish swim distance relative to mean swim distance of fishtreated with 10 mM PTZ alone. Data points represent individualzebrafish; bars shown mean±SD for each treatment group. P values at thetop of each graph show statistical significance for comparison oftreatment groups to PTZ-only group as determined by Kruskal-Wallisnonparametric ANOVA with Dunn's post-hoc correction for multiplecomparisons; p values over horizontal bars are for comparisons oftreatment groups to zebrafish untreated with PTZ or drug (****:p<0.0001; ***: p<0.001; **: p<0.01).

FIG. 6 : In vivo results for treatment of Parkinson's disease (PD) usingthe Pink1 mutant zebrafish model using NT-108. “VK2” refers to thenaturally occurring Vitamin K2.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

In some aspects, the present disclosure provides compounds which containa 1,4-napthoquinone core which may be used in the treatment of epilepsyand disorders of the mitochondria. These compounds may further comprisean additional unsaturated group such as an alkene or alkyne.

I. Definitions

When used in the context of a chemical group: “hydrogen” means —H;“hydroxy” means —OH; “oxo” means ═O; “carbonyl” means —C(═O)—; “carboxy”means —C(═O)OH (also written as —COOH or —CO₂H); “halo” meansindependently —F, —Cl, —Br or —I; “amino” means —NH₂; “hydroxyamino”means —NHOH; “nitro” means —NO₂; imino means ═NH; “cyano” means —CN;“isocyanate” means —N═C═O; “azido” means —N₃; in a monovalent context“phosphate” means —OP(O)(OH)₂ or a deprotonated form thereof; in adivalent context “phosphate” means —OP(O)(OH)O— or a deprotonated formthereof; “mercapto” means —SH; and “thio” means ═S; “sulfonyl” means—S(O)₂—; “aminosulfonyl” means —S(O)₂NH₂; “sulfonyl” means —S(O)₂OH; and“sulfinyl” means —S(O)—.

In the context of chemical formulas, the symbol “—” means a single bond,“═” means a double bond, and “≡” means triple bond. The symbol “

” represents an optional bond, which if present is either single ordouble. The symbol “

” represents a single bond or a double bond. Thus, the formula

covers, for example,

And it is understood that no one such ring atom forms part of more thanone double bond. Furthermore, it is noted that the covalent bond symbol“—”, when connecting one or two stereogenic atoms, does not indicate anypreferred stereochemistry. Instead, it covers all stereoisomers as wellas mixtures thereof. The symbol “

”, when drawn perpendicularly across a bond (e.g.,

for methyl) indicates a point of attachment of the group. It is notedthat the point of attachment is typically only identified in this mannerfor larger groups in order to assist the reader in unambiguouslyidentifying a point of attachment. The symbol “

” means a single bond where the group attached to the thick end of thewedge is “out of the page.” The symbol “

” means a single bond where the group attached to the thick end of thewedge is “into the page”. The symbol “

” means a single bond where the geometry around a double bond (e.g.,either E or Z) is undefined. Both options, as well as combinationsthereof are therefore intended. Any undefined valency on an atom of astructure shown in this application implicitly represents a hydrogenatom bonded to that atom. A bold dot on a carbon atom indicates that thehydrogen attached to that carbon is oriented out of the plane of thepaper.

When a variable is depicted as a “floating group” on a ring system, forexample, the group “R” in the formula:

then the variable may replace any hydrogen atom attached to any of thering atoms, including a depicted, implied, or expressly definedhydrogen, so long as a stable structure is formed. When a variable isdepicted as a “floating group” on a fused ring system, as for examplethe group “R” in the formula:

then the variable may replace any hydrogen attached to any of the ringatoms of either of the fused rings unless specified otherwise.Replaceable hydrogens include depicted hydrogens (e.g., the hydrogenattached to the nitrogen in the formula above), implied hydrogens (e.g.,a hydrogen of the formula above that is not shown but understood to bepresent), expressly defined hydrogens, and optional hydrogens whosepresence depends on the identity of a ring atom (e.g., a hydrogenattached to group X, when X equals —CH—), so long as a stable structureis formed. In the example depicted, R may reside on either the5-membered or the 6-membered ring of the fused ring system. In theformula above, the subscript letter “y” immediately following the Renclosed in parentheses, represents a numeric variable. Unless specifiedotherwise, this variable can be 0, 1, 2, or any integer greater than 2,only limited by the maximum number of replaceable hydrogen atoms of thering or ring system.

For the chemical groups and compound classes, the number of carbon atomsin the group or class is as indicated as follows: “Cn” defines the exactnumber (n) of carbon atoms in the group/class. “C≤n” defines the maximumnumber (n) of carbon atoms that can be in the group/class, with theminimum number as small as possible for the group/class in question,e.g., it is understood that the minimum number of carbon atoms in thegroup “alkenyl_((C≤8))” or the class “alkene_((C≤8))” is two. Comparewith “alkoxy_((C≤10))”, which designates alkoxy groups having from 1 to10 carbon atoms. “Cn-n′” defines both the minimum (n) and maximum number(n′) of carbon atoms in the group. Thus, “alkyl_((C2-10))” designatesthose alkyl groups having from 2 to 10 carbon atoms. These carbon numberindicators may precede or follow the chemical groups or class itmodifies and it may or may not be enclosed in parenthesis, withoutsignifying any change in meaning. Thus, the terms “C5 olefin”,“C5-olefin”, “olefin_((C5))”, and “olefin_(C5)” are all synonymous. Whenany of the chemical groups or compound classes defined herein ismodified by the term “substituted”, any carbon atom(s) in the moietyreplacing a hydrogen atom is not counted. Thus methoxyhexyl, which has atotal of seven carbon atoms, is an example of a substitutedalkyl_((C1-6)). Unless specified otherwise, any chemical group orcompound class listed in a claim set without a carbon atom limit has acarbon atom limit of less than or equal to twelve.

The term “saturated” when used to modify a compound or chemical groupmeans the compound or chemical group has no carbon-carbon double and nocarbon-carbon triple bonds, except as noted below. When the term is usedto modify an atom, it means that the atom is not part of any double ortriple bond. In the case of substituted versions of saturated groups,one or more carbon oxygen double bond or a carbon nitrogen double bondmay be present. And when such a bond is present, then carbon-carbondouble bonds that may occur as part of keto-enol tautomerism orimine/enamine tautomerism are not precluded. When the term “saturated”is used to modify a solution of a substance, it means that no more ofthat substance can dissolve in that solution.

The term “aliphatic” when used without the “substituted” modifiersignifies that the compound or chemical group so modified is an acyclicor cyclic, but non-aromatic hydrocarbon compound or group. In aliphaticcompounds/groups, the carbon atoms can be joined together in straightchains, branched chains, or non-aromatic rings (alicyclic). Aliphaticcompounds/groups can be saturated, that is joined by singlecarbon-carbon bonds (alkanes/alkyl), or unsaturated, with one or morecarbon-carbon double bonds (alkenes/alkenyl) or with one or morecarbon-carbon triple bonds (alkynes/alkynyl).

The term “aromatic” when used to modify a compound or a chemical grouprefers to a planar unsaturated ring of atoms with 4n+2 electrons in afully conjugated cyclic 11 system.

The term “alkyl” when used without the “substituted” modifier refers toa monovalent saturated aliphatic group with a carbon atom as the pointof attachment, a linear or branched acyclic structure, and no atomsother than carbon and hydrogen. The groups —CH₃ (Me), —CH₂CH₃ (Et),—CH₂CH₂CH₃ (n-Pr or propyl), —CH(CH₃)₂ (i-Pr, ^(i)Pr or isopropyl),—CH₂CH₂CH₂CH₃ (n-Bu), —CH(CH₃)CH₂CH₃ (sec-butyl), —CH₂CH(CH₃)₂(isobutyl), —C(CH₃)₃ (tert-butyl, t-butyl, t-Bu or ^(t)Bu), and—CH₂C(CH₃)₃ (neo-pentyl) are non-limiting examples of alkyl groups. Theterm “alkanediyl” when used without the “substituted” modifier refers toa divalent saturated aliphatic group, with one or two saturated carbonatom(s) as the point(s) of attachment, a linear or branched acyclicstructure, no carbon-carbon double or triple bonds, and no atoms otherthan carbon and hydrogen. The groups —CH₂— (methylene), —CH₂CH₂—,—CH₂C(CH₃)₂CH₂—, and —CH₂CH₂CH₂— are non-limiting examples of alkanediylgroups. The term “alkylidene” when used without the “substituted”modifier refers to the divalent group ═CRR′ in which R and R′ areindependently hydrogen or alkyl. Non-limiting examples of alkylidenegroups include: ═CH₂, ═CH(CH₂CH₃), and ═C(CH₃)₂. An “alkane” refers tothe class of compounds having the formula H—R, wherein R is alkyl asthis term is defined above. When any of these terms is used with the“substituted” modifier one or more hydrogen atom has been independentlyreplaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH,—OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂,—C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃, —S(O)₂OH, or —S(O)₂NH₂.The following groups are non-limiting examples of substituted alkylgroups: —CH₂OH, —CH₂Cl, —CF₃, —CH₂CN, —CH₂C(O)OH, —CH₂C(O)OCH₃,—CH₂C(O)NH₂, —CH₂C(O)CH₃, —CH₂OCH₃, —CH₂OC(O)CH₃, —CH₂NH₂, —CH₂N(CH₃)₂,and —CH₂CH₂Cl. The term “haloalkyl” is a subset of substituted alkyl, inwhich the hydrogen atom replacement is limited to halo (i.e. —F, —Cl,—Br, or —I) such that no other atoms aside from carbon, hydrogen andhalogen are present. The group, —CH₂Cl is a non-limiting example of ahaloalkyl. The term “fluoroalkyl” is a subset of substituted alkyl, inwhich the hydrogen atom replacement is limited to fluoro such that noother atoms aside from carbon, hydrogen and fluorine are present. Thegroups —CH₂F, —CF₃, and —CH₂CF₃ are non-limiting examples of fluoroalkylgroups.

The term “cycloalkyl” when used without the “substituted” modifierrefers to a monovalent saturated aliphatic group with a carbon atom asthe point of attachment, said carbon atom forming part of one or morenon-aromatic ring structures, no carbon-carbon double or triple bonds,and no atoms other than carbon and hydrogen. Non-limiting examplesinclude: —CH(CH₂)₂ (cyclopropyl), cyclobutyl, cyclopentyl, or cyclohexyl(Cy). As used herein, the term does not preclude the presence of one ormore alkyl groups (carbon number limitation permitting) attached to acarbon atom of the non-aromatic ring structure. The term“cycloalkanediyl” when used without the “substituted” modifier refers toa divalent saturated aliphatic group with two carbon atoms as points ofattachment, no carbon-carbon double or triple bonds, and no atoms otherthan carbon and hydrogen. The group

is a non-limiting example of cycloalkanediyl group. A “cycloalkane”refers to the class of compounds having the formula H—R, wherein R iscycloalkyl as this term is defined above. When any of these terms isused with the “substituted” modifier one or more hydrogen atom has beenindependently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃,—S(O)₂OH, or —S(O)₂NH₂.

The term “alkenyl” when used without the “substituted” modifier refersto a monovalent unsaturated aliphatic group with a carbon atom as thepoint of attachment, a linear or branched, acyclic structure, at leastone nonaromatic carbon-carbon double bond, no carbon-carbon triplebonds, and no atoms other than carbon and hydrogen. Non-limitingexamples include: —CH═CH₂ (vinyl), —CH═CHCH₃, —CH═CHCH₂CH₃, —CH₂CH═CH₂(allyl), —CH₂CH═CHCH₃, and —CH═CHCH═CH₂. The term “alkenediyl” when usedwithout the “substituted” modifier refers to a divalent unsaturatedaliphatic group, with two carbon atoms as points of attachment, a linearor branched, a linear or branched acyclic structure, at least onenonaromatic carbon-carbon double bond, no carbon-carbon triple bonds,and no atoms other than carbon and hydrogen. The groups —CH═CH—,—CH═C(CH₃)CH₂—, —CH═CHCH₂—, and —CH₂CH═CHCH₂— are non-limiting examplesof alkenediyl groups. It is noted that while the alkenediyl group isaliphatic, once connected at both ends, this group is not precluded fromforming part of an aromatic structure. The terms “alkene” and “olefin”are synonymous and refer to the class of compounds having the formulaH—R, wherein R is alkenyl as this term is defined above. Similarly, theterms “terminal alkene” and “α-olefin” are synonymous and refer to analkene having just one carbon-carbon double bond, wherein that bond ispart of a vinyl group at an end of the molecule. When any of these termsare used with the “substituted” modifier one or more hydrogen atom hasbeen independently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃,—S(O)₂OH, or —S(O)₂NH₂. The groups —CH═CHF, —CH═CHCl and —CH═CHBr arenon-limiting examples of substituted alkenyl groups.

The term “alkynyl” when used without the “substituted” modifier refersto a monovalent unsaturated aliphatic group with a carbon atom as thepoint of attachment, a linear or branched acyclic structure, at leastone carbon-carbon triple bond, and no atoms other than carbon andhydrogen. As used herein, the term alkynyl does not preclude thepresence of one or more non-aromatic carbon-carbon double bonds. Thegroups —C≡CH, —C≡CCH₃, and —CH₂C≡CCH₃ are non-limiting examples ofalkynyl groups. An “alkyne” refers to the class of compounds having theformula H—R, wherein R is alkynyl. When any of these terms are used withthe “substituted” modifier one or more hydrogen atom has beenindependently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃,—S(O)₂OH, or —S(O)₂NH₂.

The term “aryl” when used without the “substituted” modifier refers to amonovalent unsaturated aromatic group with an aromatic carbon atom asthe point of attachment, said carbon atom forming part of a one or morearomatic ring structure, wherein the ring atoms are all carbon, andwherein the group consists of no atoms other than carbon and hydrogen.If more than one ring is present, the rings may be fused or unfused.Unfused rings are connected with a covalent bond. As used herein, theterm aryl does not preclude the presence of one or more alkyl groups(carbon number limitation permitting) attached to the first aromaticring or any additional aromatic ring present. Non-limiting examples ofaryl groups include phenyl (Ph), methylphenyl, (dimethyl)phenyl,—C₆H₄CH₂CH₃ (ethylphenyl), naphthyl, and a monovalent group derived frombiphenyl (e.g., 4-phenylphenyl). The term “arenediyl” when used withoutthe “substituted” modifier refers to a divalent aromatic group with twoaromatic carbon atoms as points of attachment, said carbon atoms formingpart of one or more six-membered aromatic ring structure(s) wherein thering atoms are all carbon, and wherein the monovalent group consists ofno atoms other than carbon and hydrogen. As used herein, the termarenediyl does not preclude the presence of one or more alkyl groups(carbon number limitation permitting) attached to the first aromaticring or any additional aromatic ring present. If more than one ring ispresent, the rings may be fused or unfused. Unfused rings are connectedwith a covalent bond. Non-limiting examples of arenediyl groups include:

An “arene” refers to the class of compounds having the formula H—R,wherein R is aryl as that term is defined above. Benzene and toluene arenon-limiting examples of arenes. When any of these terms are used withthe “substituted” modifier one or more hydrogen atom has beenindependently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃,—S(O)₂OH, or —S(O)₂NH₂.

The term “aralkyl” when used without the “substituted” modifier refersto the monovalent group -alkanediyl-aryl, in which the terms alkanediyland aryl are each used in a manner consistent with the definitionsprovided above. Non-limiting examples are: phenylmethyl (benzyl, Bn) and2-phenyl-ethyl. When the term aralkyl is used with the “substituted”modifier one or more hydrogen atom from the alkanediyl and/or the arylgroup has been independently replaced by —OH, —F, —Cl, —Br, —I, —NH₂,—NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃,—NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃,—NHC(O)CH₃, —S(O)₂OH, or —S(O)₂NH₂. Non-limiting examples of substitutedaralkyls are: (3-chlorophenyl)-methyl, and 2-chloro-2-phenyl-eth-1-yl.

The term “heteroaryl” when used without the “substituted” modifierrefers to a monovalent aromatic group with an aromatic carbon atom ornitrogen atom as the point of attachment, said carbon atom or nitrogenatom forming part of one or more aromatic ring structures wherein atleast one of the ring atoms is nitrogen, oxygen or sulfur, and whereinthe heteroaryl group consists of no atoms other than carbon, hydrogen,aromatic nitrogen, aromatic oxygen and aromatic sulfur. If more than onering is present, the rings may be fused or unfused. Unfused rings areconnected with a covalent bond. As used herein, the term heteroaryl doesnot preclude the presence of one or more alkyl or aryl groups (carbonnumber limitation permitting) attached to the aromatic ring or aromaticring system. Non-limiting examples of heteroaryl groups include furanyl,imidazolyl, indolyl, indazolyl (Im), isoxazolyl, methylpyridinyl,oxazolyl, phenylpyridinyl, pyridinyl (pyridyl), pyrrolyl, pyrimidinyl,pyrazinyl, quinolyl, quinazolyl, quinoxalinyl, triazinyl, tetrazolyl,thiazolyl, thienyl, and triazolyl. The term “N-heteroaryl” refers to aheteroaryl group with a nitrogen atom as the point of attachment. A“heteroarene” refers to the class of compounds having the formula H—R,wherein R is heteroaryl. Pyridine and quinoline are non-limitingexamples of heteroarenes. When these terms are used with the“substituted” modifier one or more hydrogen atom has been independentlyreplaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH,—OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂,—C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃, —S(O)₂OH, or —S(O)₂NH₂.

The term “heterocycloalkyl” when used without the “substituted” modifierrefers to a monovalent non-aromatic group with a carbon atom or nitrogenatom as the point of attachment, said carbon atom or nitrogen atomforming part of one or more non-aromatic ring structures wherein atleast one of the ring atoms is nitrogen, oxygen or sulfur, and whereinthe heterocycloalkyl group consists of no atoms other than carbon,hydrogen, nitrogen, oxygen and sulfur. If more than one ring is present,the rings may be fused or unfused. As used herein, the term does notpreclude the presence of one or more alkyl groups (carbon numberlimitation permitting) attached to the ring or ring system. Also, theterm does not preclude the presence of one or more double bonds in thering or ring system, provided that the resulting group remainsnon-aromatic. Non-limiting examples of heterocycloalkyl groups includeaziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl,morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydrothiofuranyl,tetrahydropyranyl, pyranyl, oxiranyl, and oxetanyl. The term“N-heterocycloalkyl” refers to a heterocycloalkyl group with a nitrogenatom as the point of attachment. N-pyrrolidinyl is an example of such agroup. When these terms are used with the “substituted” modifier one ormore hydrogen atom has been independently replaced by —OH, —F, —Cl, —Br,—I, —NH₂, —NO₂, —CO₂H, —CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃,—NHCH₃, —NHCH₂CH₃, —N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂,—OC(O)CH₃, —NHC(O)CH₃, —S(O)₂OH, or —S(O)₂NH₂.

The term “acyl” when used without the “substituted” modifier refers tothe group —C(O)R, in which R is a hydrogen, alkyl, cycloalkyl, or arylas those terms are defined above. The groups, —CHO, —C(O)CH₃ (acetyl,Ac), —C(O)CH₂CH₃, —C(O)CH(CH₃)₂, —C(O)CH(CH₂)₂, —C(O)C₆H₅, and—C(O)C₆H₄CH₃ are non-limiting examples of acyl groups. A “thioacyl” isdefined in an analogous manner, except that the oxygen atom of the group—C(O)R has been replaced with a sulfur atom, —C(S)R. The term “aldehyde”corresponds to an alkyl group, as defined above, attached to a —CHOgroup. When any of these terms are used with the “substituted” modifierone or more hydrogen atom (including a hydrogen atom directly attachedto the carbon atom of the carbonyl or thiocarbonyl group, if any) hasbeen independently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃,—S(O)₂OH, or —S(O)₂NH₂. The groups, —C(O)CH₂CF₃, —CO₂H (carboxyl),—CO₂CH₃ (methylcarboxyl), —CO₂CH₂CH₃, —C(O)NH₂ (carbamoyl), and—CON(CH₃)₂, are non-limiting examples of substituted acyl groups.

The term “alkoxy” when used without the “substituted” modifier refers tothe group —OR, in which R is an alkyl, as that term is defined above.Non-limiting examples include: —OCH₃ (methoxy), —OCH₂CH₃ (ethoxy),—OCH₂CH₂CH₃, —OCH(CH₃)₂ (isopropoxy), —OC(CH₃)₃ (tert-butoxy),—OCH(CH₂)₂, —O-cyclopentyl, and —O-cyclohexyl. The terms “cycloalkoxy”,“alkenyloxy”, “alkynyloxy”, “aryloxy”, “aralkoxy”, “heteroaryloxy”,“heterocycloalkoxy”, and “acyloxy”, when used without the “substituted”modifier, refers to groups, defined as —OR, in which R is cycloalkyl,alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heterocycloalkyl, and acyl,respectively. The term “alkylthio” and “acylthio” when used without the“substituted” modifier refers to the group —SR, in which R is an alkyland acyl, respectively. The term “alcohol” corresponds to an alkane, asdefined above, wherein at least one of the hydrogen atoms has beenreplaced with a hydroxy group. The term “ether” corresponds to analkane, as defined above, wherein at least one of the hydrogen atoms hasbeen replaced with an alkoxy group. When any of these terms is used withthe “substituted” modifier one or more hydrogen atom has beenindependently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃,—S(O)₂OH, or —S(O)₂NH₂.

The term “alkylamino” when used without the “substituted” modifierrefers to the group —NHR, in which R is an alkyl, as that term isdefined above. Non-limiting examples include: —NHCH₃ and —NHCH₂CH₃. Theterm “dialkylamino” when used without the “substituted” modifier refersto the group —NRR′, in which R and R′ can be the same or different alkylgroups, or R and R′ can be taken together to represent an alkanediyl.Non-limiting examples of dialkylamino groups include: —N(CH₃)₂ and—N(CH₃)(CH₂CH₃). The terms “cycloalkylamino”, “alkenylamino”,“alkynylamino”, “arylamino”, “aralkylamino”, “heteroarylamino”,“heterocycloalkylamino”, “alkoxyamino”, and “alkylsulfonylamino” whenused without the “substituted” modifier, refers to groups, defined as—NHR, in which R is cycloalkyl, alkenyl, alkynyl, aryl, aralkyl,heteroaryl, heterocycloalkyl, alkoxy, and alkylsulfonyl, respectively. Anon-limiting example of an arylamino group is —NHC₆H₅. The term “amido”(acylamino), when used without the “substituted” modifier, refers to thegroup —NHR, in which R is acyl, as that term is defined above. Anon-limiting example of an amido group is —NHC(O)CH₃. The term“alkylimino” when used without the “substituted” modifier refers to thedivalent group ═NR, in which R is an alkyl, as that term is definedabove. When any of these terms is used with the “substituted” modifierone or more hydrogen atom attached to a carbon atom has beenindependently replaced by —OH, —F, —Cl, —Br, —I, —NH₂, —NO₂, —CO₂H,—CO₂CH₃, —CN, —SH, —OCH₃, —OCH₂CH₃, —C(O)CH₃, —NHCH₃, —NHCH₂CH₃,—N(CH₃)₂, —C(O)NH₂, —C(O)NHCH₃, —C(O)N(CH₃)₂, —OC(O)CH₃, —NHC(O)CH₃,—S(O)₂OH, or —S(O)₂NH₂. The groups —NHC(O)OCH₃ and —NHC(O)NHCH₃ arenon-limiting examples of substituted amido groups.

An “active ingredient” (AI) (also referred to as an active compound,active substance, active agent, pharmaceutical agent, agent,biologically active molecule, or a therapeutic compound) is theingredient in a pharmaceutical drug or a pesticide that is biologicallyactive. The similar terms active pharmaceutical ingredient (API) andbulk active are also used in medicine, and the term active substance maybe used for pesticide formulations.

The terms “comprise,” “have” and “include” are open-ended linking verbs.Any forms or tenses of one or more of these verbs, such as “comprises,”“comprising,” “has,” “having,” “includes” and “including,” are alsoopen-ended. For example, any method that “comprises,” “has” or“includes” one or more steps is not limited to possessing only those oneor more steps and also covers other unlisted steps.

The term “effective,” as that term is used in the specification and/orclaims, means adequate to accomplish a desired, expected, or intendedresult. “Effective amount,” “Therapeutically effective amount” or“pharmaceutically effective amount” when used in the context of treatinga patient or subject with a compound means that amount of the compoundwhich, when administered to a subject or patient for treating orpreventing a disease, is an amount sufficient to effect such treatmentor prevention of the disease.

An “excipient” is a pharmaceutically acceptable substance formulatedalong with the active ingredient(s) of a medication, pharmaceuticalcomposition, formulation, or drug delivery system. Excipients may beused, for example, to stabilize the composition, to bulk up thecomposition (thus often referred to as “bulking agents,” “fillers,” or“diluents” when used for this purpose), or to confer a therapeuticenhancement on the active ingredient in the final dosage form, such asfacilitating drug absorption, reducing viscosity, or enhancingsolubility. Excipients include pharmaceutically acceptable versions ofantiadherents, binders, coatings, colors, disintegrants, flavors,glidants, lubricants, preservatives, sorbents, sweeteners, and vehicles.The main excipient that serves as a medium for conveying the activeingredient is usually called the vehicle. Excipients may also be used inthe manufacturing process, for example, to aid in the handling of theactive substance, such as by facilitating powder flowability ornon-stick properties, in addition to aiding in vitro stability such asprevention of denaturation or aggregation over the expected shelf life.The suitability of an excipient will typically vary depending on theroute of administration, the dosage form, the active ingredient, as wellas other factors.

The term “hydrate” when used as a modifier to a compound means that thecompound has less than one (e.g., hemihydrate), one (e.g., monohydrate),or more than one (e.g., dihydrate) water molecules associated with eachcompound molecule, such as in solid forms of the compound.

As used herein, the term “IC₅₀” refers to an inhibitory dose which is50% of the maximum response obtained. This quantitative measureindicates how much of a particular drug or other substance (inhibitor)is needed to inhibit a given biological, biochemical or chemical process(or component of a process, i.e. an enzyme, cell, cell receptor ormicroorganism) by half.

An “isomer” of a first compound is a separate compound in which eachmolecule contains the same constituent atoms as the first compound, butwhere the configuration of those atoms in three dimensions differs.

As used herein, the term “patient” or “subject” refers to a livingmammalian organism, such as a human, monkey, cow, sheep, goat, dog, cat,mouse, rat, guinea pig, or transgenic species thereof. In certainembodiments, the patient or subject is a primate. Non-limiting examplesof human patients are adults, juveniles, infants and fetuses.

As generally used herein “pharmaceutically acceptable” refers to thosecompounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues, organs, and/or bodily fluids of human beings andanimals without excessive toxicity, irritation, allergic response, orother problems or complications commensurate with a reasonablebenefit/risk ratio.

“Pharmaceutically acceptable salts” means salts of compounds of thepresent invention which are pharmaceutically acceptable, as definedabove, and which possess the desired pharmacological activity. Suchsalts include acid addition salts formed with inorganic acids such ashydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like; or with organic acids such as1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid,2-naphthalenesulfonic acid, 3-phenylpropionic acid,4,4′-methylenebis(3-hydroxy-2-ene-1-carboxylic acid),4-methylbicyclo[2.2.2]oct-2-ene-1-carboxylic acid, acetic acid,aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acids,aromatic sulfuric acids, benzenesulfonic acid, benzoic acid,camphorsulfonic acid, carbonic acid, cinnamic acid, citric acid,cyclopentanepropionic acid, ethanesulfonic acid, fumaric acid,glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid,heptanoic acid, hexanoic acid, hydroxynaphthoic acid, lactic acid,laurylsulfuric acid, maleic acid, malic acid, malonic acid, mandelicacid, methanesulfonic acid, muconic acid, o-(4-hydroxybenzoyl)benzoicacid, oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substitutedalkanoic acids, propionic acid, p-toluenesulfonic acid, pyruvic acid,salicylic acid, stearic acid, succinic acid, tartaric acid,tertiarybutylacetic acid, trimethylacetic acid, and the like.Pharmaceutically acceptable salts also include base addition salts whichmay be formed when acidic protons present are capable of reacting withinorganic or organic bases. Acceptable inorganic bases include sodiumhydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide andcalcium hydroxide. Acceptable organic bases include ethanolamine,diethanolamine, triethanolamine, tromethamine, N-methylglucamine and thelike. It should be recognized that the particular anion or cationforming a part of any salt of this invention is not critical, so long asthe salt, as a whole, is pharmacologically acceptable. Additionalexamples of pharmaceutically acceptable salts and their methods ofpreparation and use are presented in Handbook of Pharmaceutical Salts:Properties, and Use (P. H. Stahl & C. G. Wermuth eds., Verlag HelveticaChimica Acta, 2002).

A “pharmaceutically acceptable carrier,” “drug carrier,” or simply“carrier” is a pharmaceutically acceptable substance formulated alongwith the active ingredient medication that is involved in carrying,delivering and/or transporting a chemical agent. Drug carriers may beused to improve the delivery and the effectiveness of drugs, includingfor example, controlled-release technology to modulate drugbioavailability, decrease drug metabolism, and/or reduce drug toxicity.Some drug carriers may increase the effectiveness of drug delivery tothe specific target sites. Examples of carriers include: liposomes,microspheres (e.g., made of poly(lactic-co-glycolic) acid), albuminmicrospheres, synthetic polymers, nanofibers, protein-DNA complexes,protein conjugates, erythrocytes, virosomes, and dendrimers.

A “pharmaceutical drug” (also referred to as a pharmaceutical,pharmaceutical agent, pharmaceutical preparation, pharmaceuticalcomposition, pharmaceutical formulation, pharmaceutical product,medicinal product, medicine, medication, medicament, or simply a drug)is a drug used to diagnose, cure, treat, or prevent disease. An activeingredient (AI) (defined above) is the ingredient in a pharmaceuticaldrug or a pesticide that is biologically active. The similar termsactive pharmaceutical ingredient (API) and bulk active are also used inmedicine, and the term active substance may be used for pesticideformulations. Some medications and pesticide products may contain morethan one active ingredient. In contrast with the active ingredients, theinactive ingredients are usually called excipients (defined above) inpharmaceutical contexts.

“Prevention” or “preventing” includes: (1) inhibiting the onset of adisease in a subject or patient which may be at risk and/or predisposedto the disease but does not yet experience or display any or all of thepathology or symptomatology of the disease, and/or (2) slowing the onsetof the pathology or symptomatology of a disease in a subject or patientwhich may be at risk and/or predisposed to the disease but does not yetexperience or display any or all of the pathology or symptomatology ofthe disease.

A “stereoisomer” or “optical isomer” is an isomer of a given compound inwhich the same atoms are bonded to the same other atoms, but where theconfiguration of those atoms in three dimensions differs. “Enantiomers”are stereoisomers of a given compound that are mirror images of eachother, like left and right hands. “Diastereomers” are stereoisomers of agiven compound that are not enantiomers. Chiral molecules contain achiral center, also referred to as a stereocenter or stereogenic center,which is any point, though not necessarily an atom, in a moleculebearing groups such that an interchanging of any two groups leads to astereoisomer. In organic compounds, the chiral center is typically acarbon, phosphorus or sulfur atom, though it is also possible for otheratoms to be stereocenters in organic and inorganic compounds. A moleculecan have multiple stereocenters, giving it many stereoisomers. Incompounds whose stereoisomerism is due to tetrahedral stereogeniccenters (e.g., tetrahedral carbon), the total number of hypotheticallypossible stereoisomers will not exceed 2^(n), where n is the number oftetrahedral stereocenters. Molecules with symmetry frequently have fewerthan the maximum possible number of stereoisomers. A 50:50 mixture ofenantiomers is referred to as a racemic mixture. Alternatively, amixture of enantiomers can be enantiomerically enriched so that oneenantiomer is present in an amount greater than 50%. Typically,enantiomers and/or diastereomers can be resolved or separated usingtechniques known in the art. It is contemplated that that for anystereocenter or axis of chirality for which stereochemistry has not beendefined, that stereocenter or axis of chirality can be present in its Rform, S form, or as a mixture of the R and S forms, including racemicand non-racemic mixtures. As used herein, the phrase “substantially freefrom other stereoisomers” means that the composition contains ≤15%, morepreferably ≤10%, even more preferably ≤5%, or most preferably ≤1% ofanother stereoisomer(s).

“Treatment” or “treating” includes (1) inhibiting a disease in a subjector patient experiencing or displaying the pathology or symptomatology ofthe disease (e.g., arresting further development of the pathology and/orsymptomatology), (2) ameliorating a disease in a subject or patient thatis experiencing or displaying the pathology or symptomatology of thedisease (e.g., reversing the pathology and/or symptomatology), and/or(3) effecting any measurable decrease in a disease in a subject orpatient that is experiencing or displaying the pathology orsymptomatology of the disease.

The above definitions supersede any conflicting definition in anyreference that is incorporated by reference herein. The fact thatcertain terms are defined, however, should not be considered asindicative that any term that is undefined is indefinite. Rather, allterms used are believed to describe the invention in terms such that oneof ordinary skill can appreciate the scope and practice the presentinvention.

II. Compounds of the Present Invention

The compounds of the present invention (also referred to as “compoundsof the present disclosure”) are shown, for example, above, in thesummary of the invention section, and in the claims below. They may bemade using the synthetic methods outlined in the Examples section. Thesemethods can be further modified and optimized using the principles andtechniques of organic chemistry as applied by a person skilled in theart. Such principles and techniques are taught, for example, in Smith,March's Advanced Organic Chemistry: Reactions, Mechanisms, andStructure, (2013), which is incorporated by reference herein. Inaddition, the synthetic methods may be further modified and optimizedfor preparative, pilot- or large-scale production, either batch ofcontinuous, using the principles and techniques of process chemistry asapplied by a person skilled in the art. Such principles and techniquesare taught, for example, in Anderson, Practical Process Research &Development—A Guide for Organic Chemists (2012), which is incorporatedby reference herein.

In some aspects, compounds are provided herein having the structure:

wherein:

-   -   X₁ and X₂ are each independently oxo or hydroxy;    -   R₂ is hydrogen, C₁₋₆ alkyl, C₆₋₁₂ aryl, C₅₋₁₂ heteroaryl, C₁₋₆        alkanediyl-C₆₋₁₂ aryl, —NH—CO—C₆₋₁₂ aryl,        —C₁₋₄alkanediyl-O—C₆₋₁₂ aryl, halogen, or a substituted version        thereof;    -   R₃ is hydrogen, amino, cyano, halo, hydroxy, nitro,        aminosulfonyl, hydroxysulfonyl, C₁₋₆ alkyl, C₁₋₆ acyl, C₁₋₆        alkoxy, C₁₋₆ alkylamino, C₁₋₈ dialkylamino, or a substituted        version of the last five groups;    -   Y is —NH— or —O—;    -   wherein when Y is —NH— and R₂ is hydrogen,        -   R₁ is C₆₋₁₈ alkenyl; —(CH₂)_(x)C≡CR_(a); wherein: x is 1, 2,            3, or 4 and R_(a) is C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈ alkynyl,            C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted version of            any of these groups;        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,                substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, or 2; or        -   a group of the formula:

-   -   -   wherein:            -   z is 1, 2, or 3            -   R_(e) is halo, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆                alkylsulfonylamino, or a substituted version of any of                the last three groups; or        -   a group of the formula:

-   -   -   wherein:            -   a is 1, 2, or 3            -   R_(f) is C₆₋₁₂ aryl, C₇₋₁₂ aralkyl, or a substituted                version of any of either groups;

    -   wherein when Y is —NH— and R₂ is not hydrogen; then:        -   R₁ is C₆₋₁₂ alkenyl, C₆₋₁₂ alkynyl, C₆₋₁₂ aryl, C₁₋₁₂            heteroaryl, or a substituted version of any of these groups,            or —Y′—X₃—R_(g), wherein:            -   Y′ is C₁₋₆ alkynediyl, C₆₋₁₂ arenediyl, or a substituted                version of either group;            -   X₃ is a covalent bond, —O—, —NHC(O)—, or —C(O)NH—; and            -   R_(g) is C₁₋₆ alkyl C₁₋₈ alkenyl, C₁₋₈ alkynyl, C₆₋₁₂                aryl, C₁₋₁₂ heteroaryl, C₇₋₁₂ aralkyl, or a substituted                version of either group; or

    -   wherein when Y is —O—,        -   R₁ is C₁₋₁₈ alkyl, substituted C₁₋₁₈ alkyl, C₁₋₁₈ alkenyl,            substituted C₁₋₁₈ alkenyl, C₁₋₁₈ alkynyl, and substituted            C₁₋₁₈ alkynyl;        -   a group of the formula:

-   -   -   wherein:            -   R_(b) is C₁₋₈ alkyl, substituted C₁₋₈ alkyl, C₆₋₁₂ aryl,                substituted C₆₋₁₂ aryl, or —C(O)R_(d);                -   wherein: R_(d) is amino, hydroxy, —NHNH₂, C₁₋₆                    alkoxy, C₁₋₆ alkylamino, or C₁₋₈ dialkylamino, or a                    substituted version of any of the last three groups;                    or            -   R_(c) is hydrogen, C₁₋₈ alkyl, C₁₋₈ alkenyl, C₁₋₈                alkynyl, C₆₋₁₂ aryl, C₁₋₁₂ heteroaryl, or a substituted                version of any of the last five groups; and            -   y is 0, 1, 2, or 3; or                or a pharmaceutically acceptable salt thereof;

Compounds described herein may display improved properties as comparedto previous compounds. For example, a compound including a1,4-naphthoquinone scaffold (a structural feature in the Vitamin Kcofactor family) was identified that protected neuronal mitochondrialhealth in vitro and demonstrated in vivo anti-seizure activity in mouseand zebrafish models: the toxicant-induced zebrafish seizure model(Baraban et al., 2005), the minimal clonic (6 Hz) mouse seizure model,and the corneal-kindled mouse model of medication-resistant epilepsy(Rahn et al., 2013). However, the compound's poor pharmacokineticprofile limits its potential as an AED (Rahn et al., 2013). In contrastand as shown in the below examples, several compounds are provided thatwith optimized anti-seizure activity in zebrafish and improvedpharmacokinetic properties in mice, that include 2-pentynylaminesubstituent or an isoamyloxy group on the 1,4-naphthoquinone scaffold.These promising results indicate that these new compounds warrantfurther development as potential AEDs.

All of the compounds of the present invention may be useful for theprevention and treatment of one or more diseases or disorders discussedherein or otherwise. In some embodiments, one or more of the compoundscharacterized or exemplified herein as an intermediate, a metabolite,and/or prodrug, may nevertheless also be useful for the prevention andtreatment of one or more diseases or disorders. As such unlessexplicitly stated to the contrary, all of the compounds of the presentinvention are deemed “active compounds” and “therapeutic compounds” thatare contemplated for use as active pharmaceutical ingredients (APIs).Actual suitability for human or veterinary use is typically determinedusing a combination of clinical trial protocols and regulatoryprocedures, such as those administered by the Food and DrugAdministration (FDA). In the United States, the FDA is responsible forprotecting the public health by assuring the safety, effectiveness,quality, and security of human and veterinary drugs, vaccines and otherbiological products, and medical devices.

In some embodiments, the compounds of the present invention have theadvantage that they may be more efficacious than, be less toxic than, belonger acting than, be more potent than, produce fewer side effectsthan, be more easily absorbed than, and/or have a better pharmacokineticprofile (e.g., higher oral bioavailability and/or lower clearance) than,and/or have other useful pharmacological, physical, or chemicalproperties over, compounds known in the prior art, whether for use inthe indications stated herein or otherwise.

Compounds of the present invention may contain one or moreasymmetrically-substituted carbon or nitrogen atoms and may be isolatedin optically active or racemic form. Thus, all chiral, diastereomeric,racemic form, epimeric form, and all geometric isomeric forms of achemical formula are intended, unless the specific stereochemistry orisomeric form is specifically indicated. Compounds may occur asracemates and racemic mixtures, single enantiomers, diastereomericmixtures and individual diastereomers. In some embodiments, a singlediastereomer is obtained. The chiral centers of the compounds of thepresent invention can have the S or the R configuration.

Chemical formulas used to represent compounds of the present inventionwill typically only show one of possibly several different tautomers.For example, many types of ketone groups are known to exist inequilibrium with corresponding enol groups. Similarly, many types ofimine groups exist in equilibrium with enamine groups. Regardless ofwhich tautomer is depicted for a given compound, and regardless of whichone is most prevalent, all tautomers of a given chemical formula areintended.

In addition, atoms making up the compounds of the present invention areintended to include all isotopic forms of such atoms. Isotopes, as usedherein, include those atoms having the same atomic number but differentmass numbers. By way of general example and without limitation, isotopesof hydrogen include tritium and deuterium, and isotopes of carboninclude ¹³C and ¹⁴C.

Compounds of the present invention may also exist in prodrug form. Sinceprodrugs are known to enhance numerous desirable qualities ofpharmaceuticals (e.g., solubility, bioavailability, manufacturing,etc.), the compounds employed in some methods of the invention may, ifdesired, be delivered in prodrug form. Thus, the invention contemplatesprodrugs of compounds of the present invention as well as methods ofdelivering prodrugs. Prodrugs of the compounds employed in the inventionmay be prepared by modifying functional groups present in the compoundin such a way that the modifications are cleaved, either in routinemanipulation or in vivo, to the parent compound. Accordingly, prodrugsinclude, for example, compounds described herein in which a hydroxy,amino, or carboxy group is bonded to any group that, when the prodrug isadministered to a subject, cleaves to form a hydroxy, amino, orcarboxylic acid, respectively.

It should be recognized that the particular anion or cation forming apart of any salt form of a compound provided herein is not critical, solong as the salt, as a whole, is pharmacologically acceptable.Additional examples of pharmaceutically acceptable salts and theirmethods of preparation and use are presented in Handbook ofPharmaceutical Salts: Properties, and Use (2002), which is incorporatedherein by reference.

It will be appreciated that many organic compounds can form complexeswith solvents in which they are reacted or from which they areprecipitated or crystallized. These complexes are known as “solvates.”Where the solvent is water, the complex is known as a “hydrate.” It willalso be appreciated that many organic compounds can exist in more thanone solid form, including crystalline and amorphous forms. All solidforms of the compounds provided herein, including any solvates thereofare within the scope of the present invention.

The above methods can be further modified and optimized for preparative,pilot- or large-scale production, either batch of continuous, using theprinciples and techniques of process chemistry as applied by a personskilled in the art. Such principles and techniques are taught, forexample, in Practical Process Research & Development (2012), which isincorporated by reference herein.

III. Treatment of Diseases

In some aspects, the compounds of the present invention may be used totreat a disease such as, e.g., a neurological disease or injury (e.g.,epilepsy, medication-resistant epilepsy, bipolar disorder or the manicphase of bipolar disorder, headaches, migraines, or a traumatic braininjury) or a mitochondrial disease (e.g., mitochondrial DNA (mtDNA)depletion syndrome (MDS), or dysfunctional mitochondrial respiratorychain disorder). In some aspects, the compound may be used to treat ametabolic disease or a mitochondrial disease in a mammalian subject,such as a human.

In some embodiments, the disease is a neurological disease such as,e.g., epilepsy, medication-resistant epilepsy, bipolar disorder or themanic phase of bipolar disorder, headaches, migraines, a traumatic braininjury, Parkinson's disease, Alzheimer's disease, Huntington's disease,Friedereich's Ataxia, or optic atrophy. As shown in the below examples,compounds provided herein displayed anti-seizure activity in zebrafishand improved pharmacokinetic properties in mice. In some embodiments,compounds provided herein may be used to treat epilepsy or as an AED.

Mitochondrial DNA depletion syndrome (also called MDS) are a group ofautosomal recessive disorders that are characterized by a significantdrop in mtDNA. The condition is often fatal in infancy and earlychildhood, and no approved therapies currently exist to treat thedisease. Particular forms of MDS include: myopathic form (typicallyinvolving mutations in the TK2 gene), encephalomyopathic form (typicallyinvolving mutations in SUCLA2 or RRM2B), the hepatopathic form(typically involving mutations in DGUOK, POLG, or MPV17), and theneurogastrointestinal form. In some embodiments, efficacy of a compounddisclosed herein may be tested using an Optic Atrophy 1 gene (OPA1)model of MDS.

Mitochondrial respiratory chain (MRC) function generally depends on thecoordinated expression of both nuclear (nDNA) and mitochondrial (mtDNA)genomes. Thus, mitochondrial diseases can for example be caused bygenetic defects in either the mitochondrial or the nuclear genome, or inthe cross-talk between the two. This impaired cross-talk can result innuclear-mitochondrial intergenomic communication disorders, which aretypically characterized by loss or instability of the mitochondrialgenome and, in turn, impaired maintenance of qualitative andquantitative mtDNA integrity. In children, most MRC disorders areassociated with nuclear gene defects rather than alterations in themtDNA itself. The MDSs are a clinically heterogeneous group of disorderswith an autosomal recessive pattern of transmission that have onset ininfancy or early childhood and are characterized by a reduced number ofcopies of mtDNA in affected tissues and organs. The MDSs include atleast four clinical presentations: hepatocerebral, myopathic,encephalomyopathic and neurogastrointestinal. MDS is further describedin, e.g., Nogueira et al. (2014).

In some embodiments, a compound of the present disclosure may be used totreat a mitochondrial disease, such as MDS, Alpers syndrome, Leigh'sDisease, autism, Parkinson's disease, Huntington's disease, Alzheimer'sdisease, amyotrophic lateral sclerosis (ALS) or Lou Gehrig's disease,optic atrophy, cardiomyopathy, muscular dystrophy, chronic fatigue,Friedreich ataxia, a progressive palsy, Charcot-Marie-Tooth disease, anacute kidney injury (AKI), a chronic kidney injury or disease, obesity,or diabetes mellitus. In some embodiments, the metabolic disease ischaracterized by a deficiency in the subject of metabolizing vitamin K₃into vitamin K₂. The mitochondrial disease may be, e.g., a mitochondrialmyopathy, Kearns-Sayre syndrome (KSS), chronic progressive externalophthalmoplegia (CPEO), diabetes mellitus and deafness (DAD), Leber'shereditary optic neuropathy (LHON), Leigh syndrome, “neuropathy, ataxia,retinitis pigmentosa, and ptosis” (NARP), “myoneurogenicgastrointestinal encephalopathy” (MNGIE), MERRF, “mitochondrialmyopathy, encephalomyopathy, lactic acidosis, or stroke-like symptoms”(MELAS). In some preferred embodiments, the mitochondrial disease isFriedreich's ataxia. Without wishing to be bound by any theory, data isprovided herein that is consistent with the idea that some of thecompounds of the present invention may affect or target mitochondrialfunction.

IV. Pharmaceutical Formulations and Routes of Administration

For the purpose of administration to a patient in need of suchtreatment, pharmaceutical formulations (also referred to as apharmaceutical preparations, pharmaceutical compositions, pharmaceuticalproducts, medicinal products, medicines, medications, or medicaments)comprise a therapeutically effective amount of a compound of the presentinvention formulated with one or more excipients and/or drug carriersappropriate to the indicated route of administration. In someembodiments, the compounds of the present invention are formulated in amanner amenable for the treatment of human and/or veterinary patients.In some embodiments, formulation comprises admixing or combining one ormore of the compounds of the present invention with one or more of thefollowing excipients: lactose, sucrose, starch powder, cellulose estersof alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesiumstearate, magnesium oxide, sodium and calcium salts of phosphoric andsulfuric acids, gelatin, acacia, sodium alginate, polyvinylpyrrolidone,and/or polyvinyl alcohol. In some embodiments, e.g., for oraladministration, the pharmaceutical formulation may be tableted orencapsulated. In some embodiments, the compounds may be dissolved orslurried in water, polyethylene glycol, propylene glycol, ethanol, cornoil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodiumchloride, and/or various buffers. Pharmaceutical formulations may besubjected to conventional pharmaceutical operations, such assterilization and/or may contain drug carriers and/or excipients such aspreservatives, stabilizers, wetting agents, emulsifiers, encapsulatingagents such as lipids, dendrimers, polymers, proteins such as albumin,or nucleic acids, and buffers, etc.

Pharmaceutical formulations may be administered by a variety of methods,e.g., orally or by injection (e.g. subcutaneous, intravenous,intraperitoneal, etc.). Depending on the route of administration, thecompounds of the present invention may be coated in a material toprotect the compound from the action of acids and other naturalconditions which may inactivate the compound. To administer the activecompound by other than parenteral administration, it may be necessary tocoat the compound with, or co-administer the compound with, a materialto prevent its inactivation. For example, the active compound may beadministered to a patient in an appropriate carrier, for example,liposomes, or a diluent. Pharmaceutically acceptable diluents includesaline and aqueous buffer solutions. Liposomes includewater-in-oil-in-water CGF emulsions as well as conventional liposomes.

The compounds of the present invention may also be administeredparenterally, intraperitoneally, intraspinally, or intracerebrally.Dispersions can be prepared in glycerol, liquid polyethylene glycols,and mixtures thereof and in oils. Under ordinary conditions of storageand use, these preparations may contain a preservative to prevent thegrowth of microorganisms.

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. The carrier can be a solvent or dispersionmedium containing, for example, water, ethanol, polyol (such as,glycerol, propylene glycol, and liquid polyethylene glycol, and thelike), suitable mixtures thereof, and vegetable oils. The properfluidity can be maintained, for example, by the use of a coating such aslecithin, by the maintenance of the required particle size in the caseof dispersion and by the use of surfactants. Prevention of the action ofmicroorganisms can be achieved by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, ascorbic acid,thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars, sodium chloride, orpolyalcohols such as mannitol and sorbitol, in the composition.Prolonged absorption of the injectable compositions can be brought aboutby including in the composition an agent which delays absorption, forexample, aluminum monostearate or gelatin.

The compounds of the present invention can be administered orally, forexample, with an inert diluent or an assimilable edible carrier. Thecompounds and other ingredients may also be enclosed in a hard or softshell gelatin capsule, compressed into tablets, or incorporated directlyinto the subject's diet. For oral therapeutic administration, thecompounds of the present invention may be incorporated with excipientsand used in the form of ingestible tablets, buccal tablets, troches,capsules, elixirs, suspensions, syrups, wafers, and the like. Thepercentage of the therapeutic compound in the compositions andpreparations may, of course, be varied. The amount of the therapeuticcompound in such pharmaceutical formulations is such that a suitabledosage will be obtained.

In some embodiments, the therapeutic compound may also be administeredtopically to the skin, eye, or mucosa. Alternatively, if local deliveryto the lungs is desired the therapeutic compound may be administered byinhalation in a dry-powder or aerosol formulation.

In some embodiments, it may be advantageous to formulate parenteralcompositions in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subjects tobe treated; each unit containing a predetermined quantity of therapeuticcompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. In someembodiments, the specification for the dosage unit forms of theinvention are dictated by and directly dependent on (a) the uniquecharacteristics of the therapeutic compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding such a therapeutic compound for the treatment ofa selected condition in a patient. In some embodiments, active compoundsare administered at a therapeutically effective dosage sufficient totreat a condition associated with a condition in a patient. For example,the efficacy of a compound can be evaluated in an animal model systemthat may be predictive of efficacy in treating the disease in a human oranother animal.

In some embodiments, the effective dose range for the therapeuticcompound can be extrapolated from effective doses determined in animalstudies for a variety of different animals. In general a humanequivalent dose (HED) in mg/kg can be calculated in accordance with thefollowing formula (see, e.g., Reagan-Shaw et al., FASEB J.,22(3):659-661, 2008, which is incorporated herein by reference):

HED (mg/kg)=Animal dose (mg/kg)×(Animal K_(m)/Human K_(m))

Use of the K_(m) factors in conversion results in more accurate HEDvalues, which are based on body surface area (BSA) rather than only onbody mass. K_(m) values for humans and various animals are well known.For example, the K_(m) for an average 60 kg human (with a BSA of 1.6 m²)is 37, whereas a 20 kg child (BSA 0.8 m²) would have a K_(m) of 25.K_(m) for some relevant animal models are also well known, including:mice K_(m) of 3 (given a weight of 0.02 kg and BSA of 0.007); hamsterK_(m) of 5 (given a weight of 0.08 kg and BSA of 0.02); rat K_(m) of 6(given a weight of 0.15 kg and BSA of 0.025) and monkey K_(m) of 12(given a weight of 3 kg and BSA of 0.24).

Precise amounts of the therapeutic composition depend on the judgment ofthe practitioner and are peculiar to each individual. Nonetheless, acalculated HED dose provides a general guide. Other factors affectingthe dose include the physical and clinical state of the patient, theroute of administration, the intended goal of treatment and the potency,stability and toxicity of the particular therapeutic formulation.

The actual dosage amount of a compound of the present disclosure orcomposition comprising a compound of the present disclosure administeredto a subject may be determined by physical and physiological factorssuch as type of animal treated, age, sex, body weight, severity ofcondition, the type of disease being treated, previous or concurrenttherapeutic interventions, idiopathy of the subject and on the route ofadministration. These factors may be determined by a skilled artisan.The practitioner responsible for administration will typically determinethe concentration of active ingredient(s) in a composition andappropriate dose(s) for the individual subject. The dosage may beadjusted by the individual physician in the event of any complication.

In some embodiments, the therapeutically effective amount typically willvary from about 0.001 mg/kg to about 1000 mg/kg, from about 0.01 mg/kgto about 750 mg/kg, from about 100 mg/kg to about 500 mg/kg, from about1 mg/kg to about 250 mg/kg, from about 10 mg/kg to about 150 mg/kg inone or more dose administrations daily, for one or several days (e.g.,depending of the mode of administration and the factors discussedabove). Other suitable dose ranges include 1 mg to 10,000 mg per day,100 mg to 10,000 mg per day, 500 mg to 10,000 mg per day, and 500 mg to1,000 mg per day. In some particular embodiments, the amount is lessthan 10,000 mg per day with a range of 750 mg to 9,000 mg per day.

In some embodiments, the amount of the active compound in thepharmaceutical formulation is from about 2 to about 75 weight percent.In some of these embodiments, the amount if from about 25 to about 60weight percent.

Single or multiple doses of the agents are contemplated. Desired timeintervals for delivery of multiple doses can be determined by one ofordinary skill in the art employing no more than routineexperimentation. As an example, subjects may be administered two dosesdaily at approximately 12 hour intervals. In some embodiments, the agentis administered once a day.

The agent(s) may be administered on a routine schedule. As used herein aroutine schedule refers to a predetermined designated period of time.The routine schedule may encompass periods of time which are identicalor which differ in length, as long as the schedule is predetermined. Forinstance, the routine schedule may involve administration twice a day,every day, every two days, every three days, every four days, every fivedays, every six days, a weekly basis, a monthly basis or any set numberof days or weeks there-between. Alternatively, the predetermined routineschedule may involve administration on a twice daily basis for the firstweek, followed by a daily basis for several months, etc. In otherembodiments, the invention provides that the agent(s) may be takenorally and that the timing of which is or is not dependent upon foodintake. Thus, for example, the agent can be taken every morning and/orevery evening, regardless of when the subject has eaten or will eat.

Pharmaceutical compositions of the present invention comprise aneffective amount of one or more compounds of the present invention oradditional agent dissolved or dispersed in a pharmaceutically acceptablecarrier. The phrases “pharmaceutical or pharmacologically acceptable”refers to molecular entities and compositions that do not produce anadverse, allergic or other untoward reaction when administered to ananimal, such as, for example, a human, as appropriate. The preparationof an pharmaceutical composition that contains at least one compound ofthe present invention or additional active ingredient will be known tothose of skill in the art in light of the present disclosure, asexemplified by Remington: The Science and Practice of Pharmacy, 21^(st)Ed. Lippincott Williams and Wilkins, 2005, incorporated herein byreference. Moreover, for animal (e.g., human) administration, it will beunderstood that preparations should meet sterility, pyrogenicity,general safety and purity standards as required by FDA Office ofBiological Standards.

As used herein, “pharmaceutically acceptable carrier” includes any andall solvents, dispersion media, coatings, surfactants, antioxidants,preservatives (e.g., antibacterial agents, antifungal agents), isotonicagents, absorption delaying agents, salts, preservatives, drugs, drugstabilizers, gels, binders, excipients, disintegration agents,lubricants, sweetening agents, flavoring agents, dyes, such likematerials and combinations thereof, as would be known to one of ordinaryskill in the art (see, for example, Remington's Pharmaceutical Sciences,18th Ed. Mack Printing Company, 1990, pp. 1289-1329, incorporated hereinby reference). Except insofar as any conventional carrier isincompatible with the active ingredient, its use in the pharmaceuticalcompositions is contemplated.

The compound of the present invention may comprise different types ofcarriers depending on whether it is to be administered in solid, liquidor aerosol form, and whether it need to be sterile for such routes ofadministration as injection. The present invention can be administeredintravenously, intradermally, transdermally, intrathecally,intraarterially, intraperitoneally, intranasally, intravaginally,intrarectally, topically, intramuscularly, subcutaneously, mucosally,orally, topically, locally, inhalation (e.g., aerosol inhalation),injection, infusion, continuous infusion, localized perfusion bathingtarget cells directly, via a catheter, via a lavage, in cremes, in lipidcompositions (e.g., liposomes), or by other method or any combination ofthe forgoing as would be known to one of ordinary skill in the art (see,for example, Remington's Pharmaceutical Sciences, 18th Ed. Mack PrintingCompany, 1990, incorporated herein by reference).

The compound of the present invention may be formulated into acomposition in a free base, neutral or salt form. Pharmaceuticallyacceptable salts, include the acid addition salts, e.g., those formedwith the free amino groups of a proteinaceous composition, or which areformed with inorganic acids such as for example, hydrochloric orphosphoric acids, or such organic acids as acetic, oxalic, tartaric ormandelic acid. Salts formed with the free carboxyl groups can also bederived from inorganic bases such as for example, sodium, potassium,ammonium, calcium or ferric hydroxides; or such organic bases asisopropylamine, trimethylamine, histidine or procaine. Upon formulation,solutions will be administered in a manner compatible with the dosageformulation and in such amount as is therapeutically effective. Theformulations are easily administered in a variety of dosage forms suchas formulated for parenteral administrations such as injectablesolutions, or aerosols for delivery to the lungs, or formulated foralimentary administrations such as drug release capsules and the like.

Further in accordance with the present invention, the composition of thepresent invention suitable for administration is provided in apharmaceutically acceptable carrier with or without an inert diluent.The carrier should be assimilable and includes liquid, semi-solid, i.e.,pastes, or solid carriers. Except insofar as any conventional media,agent, diluent or carrier is detrimental to the recipient or to thetherapeutic effectiveness of a the composition contained therein, itsuse in administrable composition for use in practicing the methods ofthe present invention is appropriate. Examples of carriers or diluentsinclude fats, oils, water, saline solutions, lipids, liposomes, resins,binders, fillers and the like, or combinations thereof. The compositionmay also comprise various antioxidants to retard oxidation of one ormore component. Additionally, the prevention of the action ofmicroorganisms can be brought about by preservatives such as variousantibacterial and antifungal agents, including but not limited toparabens (e.g., methylparabens, propylparabens), chlorobutanol, phenol,sorbic acid, thimerosal or combinations thereof.

In accordance with the present invention, the composition is combinedwith the carrier in any convenient and practical manner, i.e., bysolution, suspension, emulsification, admixture, encapsulation,absorption and the like. Such procedures are routine for those skilledin the art.

In a specific embodiment of the present invention, the composition iscombined or mixed thoroughly with a semi-solid or solid carrier. Themixing can be carried out in any convenient manner such as grinding.Stabilizing agents can be also added in the mixing process in order toprotect the composition from loss of therapeutic activity, i.e.,denaturation in the stomach. Examples of stabilizers for use in an thecomposition include buffers, amino acids such as glycine and lysine,carbohydrates such as dextrose, mannose, galactose, fructose, lactose,sucrose, maltose, sorbitol, mannitol, etc.

In further embodiments, the present invention may concern the use of apharmaceutical lipid vehicle compositions that include a compound of thepresent invention, one or more lipids, and an aqueous solvent. As usedherein, the term “lipid” will be defined to include any of a broad rangeof substances that is characteristically insoluble in water andextractable with an organic solvent. This broad class of compounds arewell known to those of skill in the art, and as the term “lipid” is usedherein, it is not limited to any particular structure. Examples includecompounds which contain long-chain aliphatic hydrocarbons and theirderivatives. A lipid may be naturally occurring or synthetic (i.e.,designed or produced by man). However, a lipid is usually a biologicalsubstance. Biological lipids are well known in the art, and include forexample, neutral fats, phospholipids, phosphoglycerides, steroids,terpenes, lysolipids, glycosphingolipids, glycolipids, sulphatides,lipids with ether and ester-linked fatty acids and polymerizable lipids,and combinations thereof. Of course, compounds other than thosespecifically described herein that are understood by one of skill in theart as lipids are also encompassed by the compositions and methods ofthe present invention.

One of ordinary skill in the art would be familiar with the range oftechniques that can be employed for dispersing a composition in a lipidvehicle. For example, the compound of the present invention may bedispersed in a solution containing a lipid, dissolved with a lipid,emulsified with a lipid, mixed with a lipid, combined with a lipid,covalently bonded to a lipid, contained as a suspension in a lipid,contained or complexed with a micelle or liposome, or otherwiseassociated with a lipid or lipid structure by any means known to thoseof ordinary skill in the art. The dispersion may or may not result inthe formation of liposomes.

The actual dosage amount of a composition of the present inventionadministered to an animal patient can be determined by physical andphysiological factors such as body weight, severity of condition, thetype of disease being treated, previous or concurrent therapeuticinterventions, idiopathy of the patient and on the route ofadministration. Depending upon the dosage and the route ofadministration, the number of administrations of a preferred dosageand/or an effective amount may vary according to the response of thesubject. The practitioner responsible for administration will, in anyevent, determine the concentration of active ingredient(s) in acomposition and appropriate dose(s) for the individual subject.

A. Alimentary Compositions and Formulations

In preferred embodiments of the present invention, the compound of thepresent invention are formulated to be administered via an alimentaryroute. Alimentary routes include all possible routes of administrationin which the composition is in direct contact with the alimentary tract.Specifically, the pharmaceutical compositions disclosed herein may beadministered orally, buccally, rectally, or sublingually. As such, thesecompositions may be formulated with an inert diluent or with anassimilable edible carrier, or they may be enclosed in hard- orsoft-shell gelatin capsule, or they may be compressed into tablets, orthey may be incorporated directly with the food of the diet.

In certain embodiments, the active compounds may be incorporated withexcipients and used in the form of ingestible tablets, buccal tables,troches, capsules, elixirs, suspensions, syrups, wafers, and the like(Mathiowitz et al., 1997; Hwang et al., 1998; U.S. Pat. Nos. 5,641,515;5,580,579 and 5,792,451, each specifically incorporated herein byreference in its entirety). The tablets, troches, pills, capsules andthe like may also contain the following: a binder, such as, for example,gum tragacanth, acacia, cornstarch, gelatin or combinations thereof; anexcipient, such as, for example, dicalcium phosphate, mannitol, lactose,starch, magnesium stearate, sodium saccharine, cellulose, magnesiumcarbonate or combinations thereof; a disintegrating agent, such as, forexample, corn starch, potato starch, alginic acid or combinationsthereof; a lubricant, such as, for example, magnesium stearate; asweetening agent, such as, for example, sucrose, lactose, saccharin orcombinations thereof; a flavoring agent, such as, for examplepeppermint, oil of wintergreen, cherry flavoring, orange flavoring, etc.When the dosage unit form is a capsule, it may contain, in addition tomaterials of the above type, a liquid carrier. Various other materialsmay be present as coatings or to otherwise modify the physical form ofthe dosage unit. For instance, tablets, pills, or capsules may be coatedwith shellac, sugar, or both. When the dosage form is a capsule, it maycontain, in addition to materials of the above type, carriers such as aliquid carrier. Gelatin capsules, tablets, or pills may be entericallycoated. Enteric coatings prevent denaturation of the composition in thestomach or upper bowel where the pH is acidic. See, e.g., U.S. Pat. No.5,629,001. Upon reaching the small intestines, the basic pH thereindissolves the coating and permits the composition to be released andabsorbed by specialized cells, e.g., epithelial enterocytes and Peyer'spatch M cells. A syrup of elixir may contain the active compound sucroseas a sweetening agent methyl and propylparabens as preservatives, a dyeand flavoring, such as cherry or orange flavor. Of course, any materialused in preparing any dosage unit form should be pharmaceutically pureand substantially non-toxic in the amounts employed. In addition, theactive compounds may be incorporated into sustained-release preparationand formulations.

For oral administration the compositions of the present invention mayalternatively be incorporated with one or more excipients in the form ofa mouthwash, dentifrice, buccal tablet, oral spray, or sublingualorally-administered formulation. For example, a mouthwash may beprepared incorporating the active ingredient in the required amount inan appropriate solvent, such as a sodium borate solution (Dobell'sSolution). Alternatively, the active ingredient may be incorporated intoan oral solution such as one containing sodium borate, glycerin andpotassium bicarbonate, or dispersed in a dentifrice, or added in atherapeutically-effective amount to a composition that may includewater, binders, abrasives, flavoring agents, foaming agents, andhumectants. Alternatively the compositions may be fashioned into atablet or solution form that may be placed under the tongue or otherwisedissolved in the mouth.

Additional formulations which are suitable for other modes of alimentaryadministration include suppositories. Suppositories are solid dosageforms of various weights and shapes, usually medicated, for insertioninto the rectum. After insertion, suppositories soften, melt or dissolvein the cavity fluids. In general, for suppositories, traditionalcarriers may include, for example, polyalkylene glycols, triglyceridesor combinations thereof. In certain embodiments, suppositories may beformed from mixtures containing, for example, the active ingredient inthe range of about 0.5% to about 10%, and preferably about 1% to about2%.

B. Parenteral Compositions and Formulations

In further embodiments, a compound of the present invention may beadministered via a parenteral route. As used herein, the term“parenteral” includes routes that bypass the alimentary tract.Specifically, the pharmaceutical compositions disclosed herein may beadministered for example, but not limited to intravenously,intradermally, intramuscularly, intraarterially, intrathecally,subcutaneous, or intraperitoneally U.S. Pat. Nos. 6,613,308, 5,466,468,5,543,158; 5,641,515; and 5,399,363 (each specifically incorporatedherein by reference in its entirety).

Solutions of the active compounds as free base or pharmacologicallyacceptable salts may be prepared in water suitably mixed with asurfactant, such as hydroxypropylcellulose. Dispersions may also beprepared in glycerol, liquid polyethylene glycols, and mixtures thereofand in oils. Under ordinary conditions of storage and use, thesepreparations contain a preservative to prevent the growth ofmicroorganisms. The pharmaceutical forms suitable for injectable useinclude sterile aqueous solutions or dispersions and sterile powders forthe extemporaneous preparation of sterile injectable solutions ordispersions (U.S. Pat. No. 5,466,468, specifically incorporated hereinby reference in its entirety). In all cases the form must be sterile andmust be fluid to the extent that easy injectability exists. It must bestable under the conditions of manufacture and storage and must bepreserved against the contaminating action of microorganisms, such asbacteria and fungi. The carrier can be a solvent or dispersion mediumcontaining, for example, water, ethanol, polyol (i.e., glycerol,propylene glycol, and liquid polyethylene glycol, and the like),suitable mixtures thereof, and/or vegetable oils. Proper fluidity may bemaintained, for example, by the use of a coating, such as lecithin, bythe maintenance of the required particle size in the case of dispersionand by the use of surfactants. The prevention of the action ofmicroorganisms can be brought about by various antibacterial andantifungal agents, for example, parabens, chlorobutanol, phenol, sorbicacid, thimerosal, and the like. In many cases, it will be preferable toinclude isotonic agents, for example, sugars or sodium chloride.Prolonged absorption of the injectable compositions can be brought aboutby the use in the compositions of agents delaying absorption, forexample, aluminum monostearate and gelatin.

For parenteral administration in an aqueous solution, for example, thesolution should be suitably buffered if necessary and the liquid diluentfirst rendered isotonic with sufficient saline or glucose. Theseparticular aqueous solutions are especially suitable for intravenous,intramuscular, subcutaneous, and intraperitoneal administration. In thisconnection, sterile aqueous media that can be employed will be known tothose of skill in the art in light of the present disclosure. Forexample, one dosage may be dissolved in isotonic NaCl solution andeither added hypodermoclysis fluid or injected at the proposed site ofinfusion, (see for example, “Remington's Pharmaceutical Sciences” 15thEdition, pages 1035-1038 and 1570-1580, 1975). Some variation in dosagewill necessarily occur depending on the condition of the subject beingtreated. The person responsible for administration will, in any event,determine the appropriate dose for the individual subject. Moreover, forhuman administration, preparations should meet sterility, pyrogenicity,general safety and purity standards as required by FDA Office ofBiologics standards.

Sterile injectable solutions are prepared by incorporating the activecompounds in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the case of sterilepowders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum-drying and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof. A powdered composition is combined with a liquidcarrier such as, e.g., water or a saline solution, with or without astabilizing agent.

C. Miscellaneous Pharmaceutical Compositions and Formulations

In other preferred embodiments of the invention, the active compound maybe formulated for administration via various miscellaneous routes, forexample, topical or transdermal administration, mucosal administration(intranasal, vaginal, etc.) and/or inhalation.

Pharmaceutical compositions for topical administration may include theactive compound formulated for a medicated application such as anointment, paste, cream or powder. Ointments include all oleaginous,adsorption, emulsion and water-solubly based compositions for topicalapplication, while creams and lotions are those compositions thatinclude an emulsion base only. Topically administered medications maycontain a penetration enhancer to facilitate adsorption of the activeingredients through the skin. Suitable penetration enhancers includeglycerin, alcohols, alkyl methyl sulfoxides, pyrrolidones andluarocapram. Possible bases for compositions for topical applicationinclude polyethylene glycol, lanolin, cold cream and petrolatum as wellas any other suitable absorption, emulsion or water-soluble ointmentbase. Topical preparations may also include emulsifiers, gelling agents,and antimicrobial preservatives as necessary to preserve the activeingredient and provide for a homogenous mixture. Transdermaladministration of the present invention may also comprise the use of a“patch”. For example, the patch may supply one or more active substancesat a predetermined rate and in a continuous manner over a fixed periodof time.

In certain embodiments, the pharmaceutical compositions may be deliveredby eye drops, intranasal sprays, inhalation, and/or other aerosoldelivery vehicles. Methods for delivering compositions directly to thelungs via nasal aerosol sprays has been described e.g., in U.S. Pat.Nos. 5,756,353 and 5,804,212 (each specifically incorporated herein byreference in its entirety). Likewise, the delivery of drugs usingintranasal microparticle resins (Takenaga et al., 1998) andlysophosphatidyl-glycerol compounds (U.S. Pat. No. 5,725,871,specifically incorporated herein by reference in its entirety) are alsowell-known in the pharmaceutical arts. Likewise, transmucosal drugdelivery in the form of a polytetrafluoroetheylene support matrix isdescribed in U.S. Pat. No. 5,780,045 (specifically incorporated hereinby reference in its entirety).

The term aerosol refers to a colloidal system of finely divided solid ofliquid particles dispersed in a liquefied or pressurized gas propellant.The typical aerosol of the present invention for inhalation will consistof a suspension of active ingredients in liquid propellant or a mixtureof liquid propellant and a suitable solvent. Suitable propellantsinclude hydrocarbons and hydrocarbon ethers. Suitable containers willvary according to the pressure requirements of the propellant.Administration of the aerosol will vary according to subject's age,weight and the severity and response of the symptoms.

IV. Examples

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples which follow representtechniques discovered by the inventor to function well in the practiceof the invention, and thus can be considered to constitute preferredmodes for its practice. However, those of skill in the art should, inlight of the present disclosure, appreciate that many changes can bemade in the specific embodiments which are disclosed and still obtain alike or similar result without departing from the spirit and scope ofthe invention.

Example 1 Development of Vitamin K Analog 2-amino- and2-alkoxy-1,4-napthoquinone Compounds as Potential Therapeutics forMedication-Resistant Epilepsy

A. Results

i. Study Rationale and Medicinal Chemistry.

Previous studies of synthetic VK analogs identified several highlyneuroprotective 1,4-naphthoquinone derivatives with substituted aminogroups attached at the 2-position. In vivo screening of these compoundsin the PTZ-induced zebrafish seizure model identified thealkyne-containing derivative, 1 (Table 1), as possessing highly potentanti-seizure activity in zebrafish and mouse seizure models [Rahn](Stables et al., 2003; Stables and Kupferberg, 1997). The compoundshowed efficacy in both the 6 Hz psychomotor (minimal clonic) seizuremodel—the only acute model for medication-resistant epilepsy (Barton etal., 2001; Metcalf et al., 2017)—and the mouse corneal kindling model, achronic model for medication-resistant epilepsy (Rowley and White, 2010;Matagne and Klitgaard, 1998). In the 6 Hz model, 4/4 animals wereprotected from seizures after 0.25 hr, however protection dropped tozero by 4 hr (Rahn et al., 2014). While the compound is stable,displaying no catabolic conversion even after 2 hr in mouse serum, 1displays a short half-life of ˜1 hr when administered by intraperitonealinjection (20 mg/kg) to adult mice (FIG. 1 ). The rapid decrease inplasma concentration in vivo compared to the lack of degradation inplasma ex vivo suggests either rapid hepatic metabolism or diminishedrenal reuptake. This finding is not surprising, as terminal alkynes areknown to be oxidized and excreted rapidly by the liver and kidney. Theclearance rate of 1 in mice correlates with the short protectivetimeframe in the mouse models of epilepsy.

Thus, improving 1's drug-like properties and optimizing protection inthe PTZ-induced zebrafish seizure model could lead to an AED withprolonged efficacy in vivo. FIG. 2 illustrates the rationale behind themedicinal chemistry. All of the compounds synthesized for this studymaintain the 1,4-naphthoquinone pharmacophore. The primary targets(Group 1) possessed 1's 2-propargylamino group plus an end-cappingorganic group appended to the alkyne. Without wishing to be bound by anytheory, it is believed that substituting the terminal alkyne wouldprolong in vivo half-life. A second set (Group 2) introduced structuralalterations closer to the naphthoquinone core, either with branchingalpha to the nitrogen or lengthening the spacer between nitrogen andalkyne to two carbons, or both of these modifications. The goal was toprobe effects of variations in polarity, flexibility, and molecularshape on activity. Several compounds (Group 3) incorporated anisoprenoid chain to the nitrogen, to model the hydrophobic fourisoprenoid-repeat tail found in MK4. One derivative with a 3-methylgroup, as found in MK4, was synthesized.

These 2-amino-substituted 1,4-napthoquinones were prepared by previouslypublished methods (Josey et al., 2013), or using modifications ofreported preparations (Valente et al., 2007; Tandon et al., 2004),involving addition of the appropriate amine as either free base orammonium chloride salt to an ethanolic solution of2-bromo-1,4-naphthoquinone in the presence of triethylamine. The aminereactants were obtained from commercial sources or prepared from thecorresponding commercial alcohols, which were tosylated and subjected toclassical Gabriel synthesis: The tosylate converted to a phthalimidederivative, and the amine generated by heating in the presence ofhydrazine (FIG. 3 ). Final products were isolated in modest yields(typically, 20-50% one-step yields) as yellow, orange, or deep redsolids after purification by reversed-phase chromatography on prepackedC18 silica columns. A byproduct observed in the preparation of thesecompounds was the 3-bromo derivative, as evidenced by LC-MS (with clear[M+H]⁺ peaks corresponding to the compounds, as well as high intensity[M+H+2]⁺ peaks corresponding to the ⁸¹Br-containing molecules). Thesebromo-substituted compounds likely formed via oxidation of the Michaeladdition intermediate of amine reacted with 2-bromo-1,4-napthoquinone,rather than by elimination of bromide.

A number of 2-alkoxy-1,4-naphthoquinone targets (Group 4, FIG. 2 ) werealso prepared. Replacement of the nitrogen attachment to thenaphthoquinone scaffold with oxygen alters the redox properties, andthus mitochondrial electron-carrying properties, of the naphthoquinonemoiety. Without wishing to be bound by any theory, it is believed thatredox potential should tune drug activity based on a mitochondrial modeof action. Thus, the 2-alkoxy compounds were prepared to directlyinterrogate the necessity of the amino attachment for seizuresuppression. The compounds were synthesized by reacting2-hydroxy-1,4-naphthoquinone with tosylated commercial alcohols in DMFstirred over solid K₂CO₃, which renders the compound nucleophilic. Thedark red reaction mixtures were heated at 60° C. for 2-4 hr, at whichpoint TLC indicated complete consumption of the tosylate reactant. Thereaction mixture remains deep red due to the presence of a deprotonatedside product, which forms by C-substitution as opposed to the desiredO-alkylation. Extraction with basic aqueous solution following dilutionof the reaction mixture into dichloromethane removes most of the sideproduct. The target 2-alkoxy compounds are then purified byreversed-phase chromatography and isolated as off-white to pale yellowsolids, generally in low (10-30%) yield.

ii. In Vitro Screening for Neuroprotection.

The HT22 murine hippocampal cell system was chosen as an initialphenotypic screen for neuroprotective efficacy (Andreux et al., 2013;Andreux et al., 2014; Ohlow et al., 2017; Schubert and Maher, 2012).HT22 cells exposed to high extracellular Glu (>5 μM) undergo oxytosis, anon-apoptotic programmed cell death involving glutathione (GSH)depletion followed by ROS accumulation, oxidative damage, and acytotoxic signaling cascade (Tan et al., 2001). Glu blocks the -xCcystine antiporter, causing (Tan et al., 2001) cells to accumulatesevere oxidative stress, mitochondrial depolarization, disruptedmitochondrial morphology, and Ca²⁺ influx (Tan et al., 2001; Yang etal., 2011; Cheng et al., 2013—cellular events that may occur inneurological diseases, including epilepsy. Thus, the immortalized HT22cell line is a well-established model of neuronal oxidative stress(Sagara and Schubert, 1998; Matsumoto et al., 1996; Albrecht et al.,2010; Ha and Park, 2006; van Leyen et al., 2005; Tobaben et al., 2011),and a verified model of neurodegeneration to screen for potentialneuroprotective drugs (Albrecht et al., 2010; van Leyen et al., 2008;Lewerenz et al., 2009; Lewerenz et al., 2003).

The substituted naphthoquinones were previously shown to protect HT22from oxytosis by restoring or protecting mitochondrial function,mitochondrial morphology, and dampening production of ROS via mechanismsthat do not involve GSH restoration or upregulation of oxidant clearanceenzymes (Josey et al., 2013). Consistent with that work, most of the2-amino compounds achieved protection of HT22 cells againstglutamate-induced oxytosis in the nM to μM range. Cells exposed to 10 mMGlu were treated compounds from 0.1 nM to 10 μM to detect EC50s in the10-1000 nM range, in order to rapidly identify compounds withneuroprotective potency equal to or greater than compounds screenedpreviously. Neuroprotection based on relative cell viability compared to+Glu and −Glu control cells was measured.

FIG. 4 shows dose-response curves for representative compounds of thefour structural groups studied. Among the 2-amino compounds (1-24), themost potent had aromatic alkyne capping groups (8-11, EC₅₀s: 100-300nM). Trifluoromethyl substitution in 12 and 13 slightly dampened potency(EC₅₀s: ˜500 nM), and substitution with the polar sulfonamide group in14 substantially decreased efficacy (EC₅₀: ˜2 μM). Less protective werestructurally similar analogs with alkyl-chain groups (2-6, EC₅₀s: ≥˜1μM). Data for 7 (see in vivo toxicity, below), with an amine-containingcapping group, was poorly reproducible and showed poor protection.Compounds with lengthened chains between nitrogen and alkyne(15,16,18,20,21) and/or branching of that chain (18-21) were also lesspotent, with EC₅₀s typically >2 μM. The most potent of the branchedcompounds, 19 (˜1.5 μM), retained the single carbon linker betweennitrogen and triple bond, while the least protective, 21 (˜6 μM),possessed a bulky phenyl branching group. The “MK-like” compounds 22 and24, with an isoprenoid tail attached to the nitrogen, were not as potent(EC₅₀s: ˜9.2 and 2.7 μM, respectively), though the bromine-containing 23showed a potent EC₅₀ of 133 nM, albeit with poorly shaped titrationdata.

The alkoxy compounds 25-31 showed markedly improved potency compared toall but the most potent amino compounds. EC₅₀s for the alkoxy compoundsranged from ˜300 nM (29) to ˜24 nM (25). Though fewer alkoxy than aminocompounds were prepared and screened, the data does not seem to indicatelarge effects on neuroprotection based on the nature of the O-cappinggroup. To support the hypothesis that the replacement of N with O wasthe primary factor leading to improved neuroprotection, compound 17 wasprepared: an amine-substituted naphthoquinone with the same isoamylsubstituent as 25. This amine analog of 25 displayed poor HT22 data butindicating an EC₅₀ at least >12 μM.

iii. In Vivo Toxicology in Zebrafish.

Prior to anti-seizure activity screening, compounds were tested fortoxicity in the animal model. Zebrafish, like rodents, possess homologsfor ˜82-84% of genes associated with human disease (Howe et al., 2013;Schriml et al., 2003), and are good models for comparative mitochondrialtoxicology (Broughton et al., 2001; Artuso et al., 2012). Goodconcordance between mammalian and zebrafish models has been verified formany developed drugs (Nadanaciva et al., 2013). The route of drugexposure (absorption from media) precludes direct correlations betweenzebrafish toxicology and patient ADME profiles; nevertheless, zebrafishtoxicity provides a convenient first-pass characterization of the safetythreshold for each compound.

Zebrafish larvae, 7 days post-fertilization (dpf), were exposed tocompounds at 20, 15, 10, and 5 μM, six larvae per compound. Animals wereobserved 1, 5, and 24 hr after dosing for overt toxicity (alteredsurvival, morbidity, morphological deformity). Heart rate, behavior, andstartle response were monitored at each time point. If substantialtoxicity was observed at these concentrations, compounds weresubsequently tested at 1 and 0.1 μM. Alkyl-capped propargylaminocompounds (2-5), and uncapped 1, showed little-to-no toxicity at 5 hr oftreatment for all concentrations tested. For some of these compounds (2,5), lethal toxicity was observed at 24 hr, particularly at higherconcentrations (15-20 μM) and roughly correlated with length of thealkyl chain substituent: the shorter the chain, the more toxic. At 24hr, 1 was lethal to all fish at 20 μM, but not at lower concentrations;2 was lethal to all fish at 10 μM or above, and to 5/6 fish at 5 μM; 3was lethal above 5 μM, while 4 was only lethal to 1/6 fish at 10 μMafter 24 hr. Compound 5, with the longest (butyl) alkyl substituent onlydisplayed toxicity when dosed at 20 μM, and no toxicity at shorterexposure times. Phenyl-capped congeners displayed little toxicity. 8 wasnot lethal at any concentration; 9, 10, 12, and 13 induced only modestmorbidity (e.g., a single fish lying on its side) even at highconcentrations. Analogs with additional nitrogen-containing groups wereexceptions: 11, with a pyridyl-capped alkyne, induced convulsive swimbehavior even at short exposure times (1-5 hr), and 14, with asulfonamide group off the phenyl ring was lethal at 10 μM to all fishafter 5 hr. 7, with and additional amino group, was lethal at mostconcentrations. These three compounds were not studied further.

Longer-chain and branched-chained amino compounds exhibited moreprominent toxic effects, some even in spite of modest structuraldifferences compared to other, non-toxic compounds tested. 15 and 16,which differ from 2 and 3 only by a second carbon between the nitrogenand alkyne, were acutely lethal to most fish at the lowest concentrationafter 1 hr. Exposure to 16 was reduced to 0.1 μM to alleviate thetoxicity observed at 24 hr. As a result, synthesis of additionalstructural derivatives of this type was abandoned. The branched compound19 was toxic after 5 hr at 10 μM (1/6 fish dead) or higher. 22, with theMK-like isoprenoid tail attached to the nitrogen, displayed sedativeeffects at low concentrations and short time periods. However 23, with a3-bromo substituent, showed little toxicity.

The alkoxy-substituted naphthoquinone, 25, was representative of thealkoxy class of compounds. No acute toxicity at high concentrations wasobserved and modest toxicity (2/3 survival) at 24 hr at the highestconcentrations. No overt toxicity was observed at 10 μM or lower at anyexposure time. While some in vitro HT-22 cytotoxicity was observed forcompounds 25-31 at concentrations >10 μM, in vivo toxicity in thesetoxicological studies was not detected, or in subsequent zebrafishanti-seizure experiments.

iv. In Vivo Efficacy Studies in a Zebrafish Seizure Model

The zebrafish PTZ-induced seizure model was used to determine the mostpromising compounds' in vivo efficacy. Zebrafish larvae (7 dpf) weretreated with the compound for 1 hr prior to inducing seizures with PTZ.As established by previous larval zebrafish anti-seizure studies (Rahnet al., 2014; Hansen et al., 2004; Baraban et al., 2005), total distancetraveled by each fish after seizure induction was measured and used as aproxy for seizure activity. Following collection of locomotor data,observation of a startle response and visual inspection for abnormalmorphology confirmed that alterations in swim activity were not causedby compound toxicity. Both 20 and 17 displayed sedative effects, sothose compounds (along with 7, 11, 14-16; see above) were excluded fromthe screening.

Table 1 summarizes the effective doses of each compound for decreasingPTZ-induced seizure activity. The most effective compounds significantlydecreased PTZ-induced seizure-like locomotor activity at low doses (5 or2.5 μM) to distances statistically indistinguishable from control (fishin untreated tank water). For 1, the lead at the outset of this study, aminimum active dose of 20 μM was identified with a significant(p=0.0053) 50% decline in locomotor activity, which was greater thanuntreated control (p=0.0004). Therefore, a minimum active dose of 10 μMwas elected as the ‘go’ criterion for identifying superior compounds interms of efficacy in reducing seizures.

Most of the branched or longer-chain amino compounds (18-20) wereineffective up to at least 10 μM. Compound 22 displayed activity at 20and 40 μM that resulted from sedation, based on decreased startleresponse; 23 and 24, also with isoprenoid chains, were ineffective.Interestingly, 21, with a phenyl branch alpha to the nitrogen atom, waseffective at 5 μM (p=0.0109), reducing PTZ-induced locomotor activity tothat of control (ns difference, p>0.99). Among 2-14 (Group 1 compounds),the structure of the alkyne capping group affected anti-seizureactivity. None of the phenyl-capped compounds tested (8-10, 12, 13)showed significant activity at 10 μM. Several compounds were tested athigher doses, to no effect: 10 was ineffective at 20 μM, 8 and 9 up to40 μM. In contrast, a number of compounds with alkyl capping groupsshowed significant anti-seizure activity. Compound 2 reduced swimactivity (p=0.0026) 89% at 40 μM to levels indistinguishable fromuntreated fish, but was not effective at 20 μM or lower. Compound 4decreased PTZ-induced locomotor activity ˜80% at 20 and 10 μM, butwithout statistical significance (p=0.0678 and 0.0847, respectively).Compound 6 showed significant activity at 10 μM (p=0.0060), reducingswim distance 85% (ns compared to control, p=0.9499); at 5 μM, seizurelocomotion was reduced ˜65%, though this activity was not significantcompared to PTZ-treatment only (p=0.2381). Of this set of compounds, 3and 5—with ethyl and butyl capping groups, respectively—demonstratedconvincing anti-seizure activity. 5 completely attenuated PTZ-inducedlocomotor activity (ns compared to control, p>0.9999 at 10 μM, p=0.1901at 5 μM) at doses as low as 5 μM. The reduction in swim activity was 84%at 5 μM (p<0.0001) and 77% at 10 μM (p=0.0069). Treatment with compound3 also induced significant locomotor reduction across a range of doses:20 μM (91% reduction, p<0.0001), 10 μM (80% reduction, p<0.0001), and 5μM (62% reduction, p=0.0002). FIG. 5 a depicts the apparentdose-response behavior of induced seizure activity to treatment with 3.

Several of the alkoxy-substituted naphthoquinones were tested (25,29-31) as well. Compound 29 elicited significant reductions inPTZ-induced swim distances at 10 μM (53%, p=0.0226) and 5 μM (60%,p=0.0016; ns compared to control). 30 had significant efficacy at 10 μM(p=0.0010; 68% reduction, ns compared to control) and at 5 μM (p=0.0064,53% reduction, ns compared to control). The 5 μM treatment elicited a53% reduction in locomotor activity (p=0.0891) compared to control (noPTZ treatment, 83% lower swim distance than PTZ-only treatment). At 10μM, 31 reduced swim distance to control levels (ns, p>0.9999) with ahighly significant (p<0.0001) difference compared to PTZ treatment only;treatment effects at 5 μM were not significant (48% decrease in swimactivity, p=0.1222). Treatment with 31 also led to higher-than-normaloccurrences of toxic effects (either lack of survival or behavioralimpairments) during PTZ-induced seizure experiments.

FIG. 5 b displays anti-seizure data for compound 25, which proved to bethe most effective compound tested: significant reduction (45%,p=0.0038) of locomotor activity at a dose of 2.5 μM, though not reducedto control (p=0.0005). The high activity and good apparent dose-responsebehavior of this compound led us to select 25, along with 3, for PKstudies.

v. In Vivo Pharmacokinetic Studies in Mice

PK studies on 3 and 25 were carried out in CD-1 mice. Three males weredosed via oral gavage needle for oral administration at 20 mg/kg (10mL/kg) or via intravenous administration at 5 mg/kg (10 mL/kg). Bloodsamples were taken at time intervals between 5 min and 24 hr from eachanimal. Animals treated with either compound displayed no abnormalclinical symptoms during the IV study. For 25, plasma concentrationsdeclined in a multiphasic manner after IV administration at 5 mg/kg witha mean initial concentration (C₀) of 1,603 ng/mL. The compound displayeda high systemic clearance (CLp) of 68.8 mL/min/kg and a highsteady-state volume of distribution (V_(ss)) of 13.2 L/kg, suggesting anextensive tissue distribution. The total systemic exposure (AUC_(inf))was low at 1,211 h*ng/mL with a moderate terminal half-life (t_(1/2)) of4.31 h. For 3, C₀ was 1,523 ng/mL. The compound displayed a high CLp of188 mL/min/kg and a high Vss of 48.6 L/kg, also suggestive of extensivetissue distribution. A low AUC_(inf) of 448 h*ng/mL was determined, witha long t_(1/2) of 14.7 hr.

Initial PO experiments encountered difficulties. A formulation of 20%DMA—40% PEG300—40% H₂O for the 20 mg/kg dosing led to labored breathingand significantly reduced activity in all animals within 30 min; allmice died between the 8 and 24 hr time points. 25 rapidly reached a highC_(max) of 2,083 ng/mL within 30 min. Plasma concentrations thendeclined with a moderate t_(1/2) of 5.56 hr. AUC_(inf) was high at11,521 hr*ng/mL with a high oral bioavailability (F) of greater than100%, suggesting some saturation of clearance mechanisms. 3 reached aC_(max) of 394 ng/mL within 15 min. Plasma concentrations then declinedwith a moderate t_(1/2) of 5.27 hr. A modest AUC_(inf) (992 h*ng/mL) andgood oral bioavailability (F, 55.3%) were determined. PO PK experimentswere repeated for 25, with oral dosing at 20 mg/kg and 10 mL/kg of asuspension with 1% Tween 80:1% carboxymethylcellulose in water. Noabnormal clinical symptoms were observed following administration ofthis formulation. The test compound rapidly reached a moderate C_(max)of 282 ng/mL within 1 hr. After that, its plasma concentrations declinedwith a moderate t_(1/2) of 3.49 hr. AUC_(inf) was moderate at 1,953hr*ng/mL, with a good oral bioavailability (F) of 40.3%.

B. Discussion

There is a critical unmet clinical need for new AEDs for patients withmedication-resistant epilepsy. Prior work identified compound 1 as apotential AED based on anti-seizure activity in three vertebrate animalseizure models. These previous studies demonstrated that the2-propargylamino-substituted 1,4-naphthoquinone was a promising scaffoldfor an AED. However, anti-seizure effects rapidly diminished within 1 hrin mouse seizure models. It is hypothesized that the short half-life(t_(1/2)=0.35 hr, 20 mg/kg i.p. injection) of 1 could at least partlyexplain the drop-off in protection over time.

In this study, the PK parameters were optimized and efficacy of theprior lead by synthesizing a series of 2-substituted naphthoquinonederivatives with capped amino or alkoxy substituents. These compoundswere screened for in vitro neuroprotection and for in vivo mitigation ofPTZ-induced seizure activity in zebrafish. Two compounds, with in vivomicromolar efficacy were selected and dose responsiveness in zebrafishfor PK studies in mice. Capping the terminal alkyne of 1 with an alkylor aryl group was intended to halt rapid in vivo drug oxidation andexcretion. Additional variations in structure led to compounds withaltered chain lengths between nitrogen and alkyne, variable group sizesand functional groups present in the alkyne-capping moiety, branching atthe amino alpha carbon, and introduction of alternate unsaturated groupsin place of alkyne (FIG. 2 ). These compounds provided a range ofdrug-like properties (e.g., lipophilicity, molecular shape, andstructural rigidity) for optimization. Derivatives with the amine groupreplaced with an ether group were also synthesized, based on thehypothesis that altering the redox properties of the naphthoquinonepharmacophore would in turn affect activity. Earlier studies, whichidentified aminonaphthoquinones as promising neuroprotectants in HT22cells, indicated that 2-hydroxynaphthoquinone as a much less potentcompound for protecting neurons from oxytosis (Josey et al., 2013).However, unlike the amino compounds, 2-hydroxynaphthoquinone has a lowpKa and likely exists in its deprotonated, anionic state to asignificant extent in vitro and in vivo (Petrova et al., 1990).Deprotonation substantially alters its reduction potential and drug-likeproperties. Capping the oxygen with an alkyl substituent preventsdeprotonation, potential tautomerization to toxic 1,2-naphthoquinone,and opens synthetic space for tuning drug-like properties. Thus, it wasinvestigated if naphthoquinone redox potential, modulated by N- vs.O-substitution, has an effect on anti-seizure activity andneuroprotection, as might be expected for a drug with a mitochondrialmodulation mode of action (Ohlow et al., 2017; Vafai et al., 2016; Wenet al., 2011; Poteet et al., 2012).

Although several of the compounds tested in this study have beenreported previously (Fei et al., 2011; Fei et al., 2010; Google Patents,assignee. Synthesis method of azepine anthraquinone 2010; Jiang et al.,2010; Jiang and Wang, 2009; Wang et al., 2014; Gornostaev et al., 2016;Novel tetracyclonaphthooxazole derivative and preparation methodthereof, 2015; Adin and Fleming, 1980; Fieser, 1926; Wang et al., 2015;Kumar et al., 2017; Ogata et al., 2016; Lien et al., 2002), none hasbeen studied as a potential AED to the inventor's knowledge. Themajority of the targets are new compounds. While the yields of thereactions were modest, the short synthetic routes and ease ofpreparation lend themselves to scaled-up synthesis.

Among the amino compounds (1-24) screened In the HT22 oxytosis assay,propargylamino compounds (1-13) were more protective in general than thecompounds with branching between nitrogen and alkyne (18-21) or evencompounds with a chain between nitrogen and alkyne lengthened by asingle carbon (15, 16). In particular, the compounds (8-13) with arylsubstitution of the alkyne were more effective neuroprotectants in theHT22 assay than the other amino compounds. An exception was 14, with ahighly polar sulfonamide moiety on the phenyl ring. The more “MK-like”compounds (22, 24) with isoprenoid groups did not show enhancedneuroprotective activity compared to other compounds tested; 22 was oneof the least effective compounds screened (EC50: ˜9 μM). The greaterrigidity (i.e., fewer rotatable bonds) of aryl groups may enhance thedrug-like properties of the compounds, or the aryl groups may allow forbetter interaction with an as-yet identified target in the cells. Butwith the EC₅₀s spanning little more than an order of magnitude withoverlaps between structural classes, the in vitro data preclude drawingbroad conclusions correlating subtle structural differences toneuroprotective efficacy. Several of the alkoxy compounds were morepotent than any of the amino compounds tested, and all were roughly asprotective as the most potent amino compounds. Without wishing to bebound by any theory, it is believed that the large change in redoxpotential that an O- vs. N-substituent imparts on the naphthoquinonering, with the alkoxy compounds predicted to have a more positivepotential including more capable of accepting electrons from cellularreducing agents or enzymes (Milton et al., 2015; Fieser and Fieser,1935). The alkoxy compounds also lack the additional hydrogen bond donorprovided by an NH (group, which may alter potency in the HT22 oxytosissystem. However, in the previous study, capping the amine was found witha second, small substituent (e.g., methyl) and thereby removing thatH-bond donor tended to show little effect on compounds' efficacy in thisassay (Josey et al., 2013).

In the zebrafish PTZ-induced seizure model, several of the compoundsthat displayed neuroprotection in HT22 failed to protect againstseizures. For example, the phenyl-capped compounds (8-13) displayedlittle anti-seizure activity. These results are consistent with theprevious studies, which demonstrated that greater efficacy in the HT22oxytosis assay does not necessarily correlate with efficacy in thezebrafish seizure model. Initial results indicated that 22, which mostclosely resembles the structure of MK4, potently reduced swim activity;however, on further investigation, this result originated from a generalsedative effect rather than specific anti-seizure activity. Thecompounds proved to be compounds similar in structure to 1 but withshort (2,3,5,6) alkyl capping groups on the alkyne, as well as severalof the alkoxy compounds, particularly 25, with a five-carbon alkyl chainattached to the oxygen. Apart from the naphthoquinone core, 25 isstructurally distinct from 1. Of these active compounds, 3, 5, 25, 29,and 3 displayed significant efficacy at doses as low as 5 μM.Interestingly, 21—with phenyl branching and an elongated chain—was alsoeffective at 5 μM; no other compound in that structural class showed anyanti-seizure activity. However, the uncapped alkyne moiety would be aliability in PK studies; thus, derivative of this compound will be afocus of future studies.

25 was the only compound to show significant anti-seizure activity at2.5 μM. In the specific case of 25, the potent in vitro neuroprotectantproved to be an effective in vivo anti-seizure compound tested. Yet 3had an in vitro neuroprotective efficacy several hundred times less thanthat of 25. The mechanisms of compound uptake from tank water mayexplain some difference between activity in zebrafish and in cells.Without wishing to be bound by any theory, some of this difference tocompounds' blood-brain barrier (BBB) permeability in vivo, but certaintyof the role of the zebrafish BBB in these studies cannot be determinedat this time. While zebrafish may begin to develop a BBB with structuraland functional similarities to that of higher-vertebrates at 3 dpf, theBBB may not be fully mature until 10 dpf or later (Watanabe et al.,2012; Xie et al., 2010; Jeong et al., 2008; Fleming et al., 2013). 7 dpflarvae were used in these experiments.

PK studies were carried out in mice treated both compounds 3 and 25intravenously and per os. As hypothesized, these two compounds displayeddramatically improved PK properties compared to 1. IV-administeredhalf-life increased from ˜20 min for 1 to nearly 15 hr for 3 on cappingthe terminal alkyne with the ethyl group, data that clearly demonstratesthe PK liability of 1's oxidizable terminal alkyne. Despite the toxicityeffects observed for PO dosing in a high DMA cosolvent solutionformulation, 25 in particular showed excellent oral systemic exposure(C_(max)=2,083 ng/mL, AUC=11,521 hr*ng/mL), sustained high oralconcentrations (C=531 ng/mL at 8 hr post-dose), and good oral terminalhalf-life (t_(1/2)=5.6 hr), with acceptable systemic clearance (C_(L)=69mL/min/kg) and volume of distribution (V_(ss)=13 L/kg), and high oralbioavailability (>100%, likely due to saturation of clearancemechanisms). Repeating the oral PK studies at 20 mg/kg in a suspensionformulation led to reduced systemic exposure and therefore oralbioavailability, as expected: C_(max) and AUC were significantlydecreased (6-7 fold) and the oral bioavailability was 40%. It is likelythat a suspension formulation would need to be used for a generaltoxicology study (GLP or non-GLP). In addition to the problems observedwith a solution formulation for these compounds, much higher oral dosesare required to reach the maximum tolerated dose in toxicology studiesin mammals. The PK parameters observed for 25 when administered in asuspension are within acceptable ranges for druggable compounds,depending upon the potency of the molecule. Thus, the efficacy of 25 isstill being tested as an orally administered drug in multiple rodentepilepsy models. Further, full metabolic stability studies, aqueoussolubility studies, P450 inhibition experiments, and determining plasmaprotein binding in mouse and human plasma are being carried out. Studiesto determine brain permeation of these compounds in mice are also beingcarried out.

TABLE 1 Substituted 1,4-naphthoquinones included in this study

Neuroprotection^(a) Anti-seizure^(b) Compound R EC₅₀ (nM) activity 1 —H694^(c) 20 μM (50%)** 2 —CH₃  1219 (1062-1398) 40 μM (89%)**, + 3—CH₂CH₃   1011 (911-1123) 5 μM (62%)*** 4

   989 (878-1115) >20 μM 5

 1569 (1461-1684) 5 μM (84%)***, + 6

 2640 (2318-3007) 10 μM (85%)**, + 7

N/A^(d) N/A^(e) 8

  162 (130-202) >40 μM 9

  216 (175-267) >40 μM 10

  215 (187-246) >20 μM 11

  290 (258-327) N/A^(e) 12

  566 (419-766) >10 μM 13

  584 (511-667) >10 μM 14

 1941 (1639-2300) N/A^(e) 15

 2911 (2589-3273) N/A^(e) 16

 5603 (4952-6340) N/A^(e) 17

N/A^(d) 5 μM (76%)**, +, # 18

 2168 (1497-3140) >10 μM 19

 2145 (1767-2605) >10 μM 20

 3691 (3283-4149) >10 μM 21

 6151 (5763-6565) 5 μM (86%)*, + 22

 9222 (9464-11240) 20 μM (96%)*, +, # 23

  133 (106-166) >10 μM 24

 2738 (2301-3259) >10 μM 25

  23.9 (17.1-32.2) 2.5 μM (450)** 26

  78.0 (51.7-118) N/A^(e) 27

  168 (151-188) N/A^(e) 28

  168 (127-222) N/A^(e) 29

307 (144-651) 5 μM (60%)**, + 30

  249 (235-263) 5 μM (53%)**, + 31

  76.1 (66.6-87.0) 10 μM (89%)***, + 32

N/A^(e) N/A^(e) ^(a)In vitro neuroprotection assessed by the HT22oxytosis assay. Cell viability was estimated by CellTiter Blue treatmentwith fluorescence measurements at 490 nm. EC₅₀ (drug concentrationprotecting 50% of cells from death) values were calculated usingGraphPad Prism log(dose)-response curve fitting, based on ≥8-pointtitrations, n ≥ 2.95% CI in parentheses. ^(b)Anti-seizure activity bythe zebrafish PTZ-induced seizure model based on distance traveled (mm)as tracked by DanioVision. Concentration given is the lowest tested doseat which the drug treatment group showed significant difference comparedto normalized PTZ treatment-only groups. In parentheses, the percentreduction in mean swim activity at that dose, compared to PTZ controlfish (100% = reduction to tank water-only control swim distance). TheKruskal-Wallis nonparametric test for one-way analysis of variance(ANOVA) followed by Dunn's Method for multiple comparisons was used tocompare groups. *p <0.05, **p <0.01, ***p <0.001. +Treatment groupstatistically indistinguishable from untreated (no PTZ) control group,p >0.05. # Swim activity reduction likely non-specific result ofsedative or other drug effect. ^(c)Previously reported. ^(d)Not tested.^(e)Results gave poor EC50 curve fits.

TABLE 2 PK Summary 1 3 25 Standard Standard Standard Parameter Meandeviation Mean deviation Mean deviation Intravenous Administrationt_(1/2) (hr) 0.349 0.128 14.7 3.78 4.31 0.46 C₀ (ng/mL) 2017 570 1523254 1603 668 C_(last) (ng/mL) 1.37 0.167 2.26 0.42 T_(last) (hr) 24 0 240 AUC_(last) (hr*ng/mL) 404 58.7 421 64.8 1197 16.9 AUC_(inf) (hr*ng/mL)404 58.2 448 69 1211 20.3 MRT_(inf) (hr) 0.282 0.0382 4.28 1.14 3.2 0.03AUC_(inf)/D (hr*mg/mL) 89.6 13.7 242 3.6 CL (mL/min/kg) 83.6 11.6 18827.3 68.8 1.14 V_(ss) (L/kg) 1.43 0.387 48.6 15.9 13.2 0.21 OralAdministration T_(1/2) (hr) 5.27 0.27 5.56 0.643 C_(max) (ng/mL) 39498.3 2083 362 T_(max) (hr) 0.139 0.096 0.333 0.144 C_(last) (ng/mL) 33.14.93 531 37.8 T_(last) (hr) 8 0 8 0 AUC_(last) (hr*ng/mL) 738 49 7360294 AUC_(inf) (hr*ng/mL) 992 59.4 11521 482 MRT_(inf) (hr) 5.82 0.537.84 0.88 C_(max)/D (mg/mL) 19.8 4.92 104 18.1 AUC_(inf)/D (hr*mg/mL)49.6 3 576 24.2 F (%) 55.3 3.31 238 9.96

Example 2

A. Methods

i. General Chemistry

Unless otherwise noted, chemicals and solvents were acquired fromcommercial sources and used as received without further purification.Yields refer to chromatographically and spectroscopically (¹H NMR)homogeneous material, unless otherwise stated. Reactions were monitoredby thin layer chromatography (TLC) carried out on precoated silica gelPE SIL G/UV plates (Whatman), using UV light to visualize spots. Flashsilica gel chromatography was carried out using prepacked silica columnson a Teledyne Isco Combiflash 200 eluting with ethyl acetate:hexane andreverse phase chromatography was performed using prepacked C18 columnsand a Teledyne Isco Combiflash 200 eluting with water:acetonitrile. Alltarget compounds were at least 95% pure, confirmed via UV detection(λ=254 nm) on a Thermo LCQ Fleet HPLC-MS using an Accucore RP-MS HPLCcolumn, 2.6 μm particle size, 30 mm×4.6 mm. Mobile phase was a gradientof water:methanol, each solvent containing 0.1% formic acid (v/v).[Agilent 1100 HPLC instrument using an ODS HYPERSIL column (5 μm, 4.6mm×250 mm) using a gradient of water/methanol with 0.1% formic acidadded.] Mass spectral data were gathered using a Thermo LCQ Fleet massspectrometer with electrospray ionization. ¹H NMR and ¹³C NMR data werecollected in deuterated solvent with a Bruker 400 MHz and referenced toresidual protio solvent or solvent carbon, respectively. Chemical shiftsare given in parts per million (δ). NMR descriptions use the followingabbreviations: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet,and br=broad peak. Coupling constants (J) reported in Hz. Chemical namesthat are followed by footnotes indicate that those compounds have beenpreviously reported.

ii. Synthesis of 2-Amino-1,4-Naphthoquinone Derivatives: GeneralProcedure

One equiv. of the appropriate amine as free base or ammonium chloridesalt was added to a solution of 2-bromo-1,4-naphthoquinone (typically,150 mg, 0.633 mmol) in absolute EtOH (20 mL) in the presence of 2.2equiv. triethylamine (194 μL, 1.39 mmol), resulting in color changesfrom yellow to deep red to brown. The reaction was stirred at roomtemperature for time periods of three hours to overnight, with reactioncompletion determined by TLC monitoring based on the absence of startingbromonaphthoquinone. Volatiles were removed by rotary evaporation andthe crude residue redissolved in 2-4 mL of solvent for purification on aTeledyne Isco Combiflash automated chromatography system usingpre-packed C18 silica gel columns and eluting with a water:acetonitrilegradient, unless otherwise indicated. Fractions monitored at 254 nm UVwere collected. Amine precursors were either commercially available freebases or ammonium chloride salts, or were prepared as ammonium chloridesalts from commercially available alcohol precursors as describedpreviously. Compound 1 had been prepared and purified as part of a priorstudy.

2-(But-2-yn-1-ylamino)naphthalene-1,4-dione (2) (Fei et al., 2011; Feiet al., 2010; Google Patents, assignee. Synthesis method of azepineanthraquinone 2010; Jiang et al., 2010; Jiang and Wang, 2009) Yield: 91mg (64%) of yellow powder. ESI-MS m/z: 226.17 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 7.98 (dd, J=7.7, 1.3 Hz, 1H), 7.95 (dd, J=7.7, 1.3 Hz, 1H),7.83 (td, J=7.5, 1.4 Hz, 1H), 7.79 (t, J=6.0 Hz, 1H), 7.74 (td, J=7.5,1.4 Hz, 1H), 5.75 (s, 1H), 3.99 (dq, J=5.0, 2.4 Hz, 2H), 1.79 (t, J=2.4Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 182.04, 181.94, 148.41, 135.32,133.35, 132.81, 130.81, 126.36, 125.84, 101.47, 80.00, 74.62, 31.92,3.53.

2-(Pent-2-yn-1-ylamino)naphthalene-1,4-dione (3) Yield: 51.6 mg (34%) ofyellow-orange powder. ESI-MS m/z: 240.10 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 7.97 (dd, J=7.7, 1.3 Hz, 1H), 7.94 (dd, J=7.6, 1.3 Hz, 1H),7.83 (td, J=7.5, 1.3 Hz, 1H), 7.78 (t, J=6.0 Hz, 1H), 7.73 (td, J=7.5,1.4 Hz, 1H), 5.75 (s, 1H), 4.00 (dt, J=5.9, 2.2 Hz, 2H), 2.18 (qt,J=7.5, 2.2 Hz, 2H), 1.04 (t, J=7.5 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ182.02, 181.94, 148.36, 135.30, 133.33, 132.79, 130.80, 126.34, 125.84,101.52, 85.68, 74.78, 31.95, 14.18, 12.10.

2-(Hex-2-yn-1-ylamino)naphthalene-1,4-dione (4) (Fei et al., 2011; Feiet al., 2010; Jiang et al., 2010) Yield: 28.8 mg (17.9%) of yellowpowder. ESI-MS m/z: 254.19 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 7.98 (dd,J=7.6, 1.3 Hz, 1H), 7.94 (dd, J=7.7, 1.3 Hz, 1H), 7.83 (td, J=7.6, 1.4Hz, 1H), 7.79 (d, J=6.0 Hz, 1H), 7.74 (td, J=7.5, 1.4 Hz, 1H), 5.77 (s,1H), 4.01 (dt, J=6.0, 2.2 Hz, 2H), 2.15 (tt, J=7.0, 2.2 Hz, 2H), 1.42(h, J=7.2 Hz, 2H), 0.90 (t, J=7.4 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ181.99, 181.97, 148.35, 135.32, 133.33, 132.80, 130.80, 126.36, 125.85,101.61, 84.19, 75.58, 31.97, 31.16, 22.02, 20.32, 13.67.

2-(Hept-2-yn-1-ylamino)naphthalene-1,4-dione (5) This synthesis wascarried out on a 176 mg (0.743 mmol) scale. Yield: 120.4 mg (60.7%) oforange-yellow powder. ESI-MS m/z: 268.25 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 7.97 (dd, J=7.7, 1.3 Hz, 1H), 7.94 (dd, J=7.7, 1.3 Hz, 1H),7.83 (td, J=7.6, 1.4 Hz, 1H), 7.78 (t, J=6.0 Hz, 1H), 7.73 (td, J=7.5,1.4 Hz, 1H), 5.76 (s, 1H), 4.01 (dt, J=6.1, 2.2 Hz, 2H), 2.17 (tt,J=6.8, 2.2 Hz, 2H), 1.45-1.25 (m, 4H), 0.82 (t, J=7.2 Hz, 3H). ¹³C NMR(101 MHz, DMSO-d₆) δ 181.97, 148.33, 135.30, 133.33, 132.78, 130.79,126.34, 125.84, 101.64, 84.31, 75.44, 31.98, 30.61, 21.73, 18.03, 13.84.

2-(Hex-5-en-2-yn-1-ylamino)naphthalene-1,4-dione (6) Synthesis of thiscompound was carried out on a 0.545 mmol scale. Yield: 24 mg (17.6%) ofyellow powder. ESI-MS m/z: 252.25 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ7.98 (dd, J=7.6, 1.3 Hz, 1H), 7.95 (dd, J=7.6, 1.3 Hz, 1H), 7.88-7.81(m, 2H), 7.74 (td, J=7.5, 1.4 Hz, 1H), 5.79 (ddd, J=22.0, 10.1, 5.1 Hz,1H), 5.79 (s, 1H), 5.28 (dq, J=17.0, 1.9 Hz, 1H), 5.07 (dq, J=10.0, 1.8Hz, 1H), 4.07 (dt, J=6.1, 2.2 Hz, 2H), 3.01 (dp, J=6.0, 2.0 Hz, 2H). ¹³CNMR (101 MHz, DMSO-d₆) δ 182.03, 181.95, 148.38, 135.34, 133.32, 133.24,132.84, 130.81, 126.38, 125.85, 116.34, 101.62, 80.75, 78.05, 31.92,31.16, 22.70.

2-((4-(Dimethylamino)but-2-yn-1-yl)amino)naphthalene-1,4-dione (7)Yield: 62.2 mg (36.3%) of yellow powder. ESI-MS m/z: 269.17 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 7.98 (dd, J=7.6, 1.3 Hz, 1H), 7.95 (dd, J=7.7,1.3 Hz, 1H), 7.89-7.79 (m, 1H), 7.74 (td, J=7.5, 1.4 Hz, 1H), 5.80 (s,1H), 4.09 (dt, J=6.2, 2.0 Hz, 2H), 3.21 (t, J=2.0 Hz, 1H), 2.13 (s, 6H).¹³C NMR (101 MHz, DMSO-d₆) δ 182.00, 181.96, 148.28, 135.33, 133.29,132.84, 130.80, 126.37, 125.86, 101.87, 80.14, 79.37, 47.60, 44.07,31.83.

2-((3-Phenylprop-2-yn-1-yl)amino)naphthalene-1,4-dione (8) (Fei et al.,2011; Fei et al., 2010; Jiang et al., 2010) This reaction was carriedout on a 1.20 mmol scale. Yield: 72.4 mg (20.9%) of orange powder.ESI-MS m/z: 288.25 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.00 (dd, J=7.7,1.3 Hz, 1H), 7.95 (ddd, J=9.7, 6.8, 3.5 Hz, 2H), 7.84 (td, J=7.5, 1.4Hz, 1H), 7.75 (td, J=7.5, 1.4 Hz, 1H), 7.47-7.31 (m, 5H), 5.89 (s, 1H),4.31 (d, J=5.9 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 182.16, 181.93,148.44, 135.32, 133.32, 132.86, 131.87, 130.86, 129.19, 129.16, 126.39,125.87, 122.39, 101.81, 85.28, 83.55, 32.40.

2-((3-(3-Fluorophenyl)prop-2-yn-1-yl)amino)naphthalene-1,4-dione (9)This reaction was carried out on a 1.25 mmol scale. Yield: 94 mg (24.6%)of bright orange powder. ESI-MS m/z: 306.17 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 7.98 (dd, J=7.6, 1.3 Hz, 1H), 7.96-7.90 (m, 2H), 7.83 (td,J=7.5, 1.3 Hz, 1H), 7.73 (td, J=7.5, 1.3 Hz, 1H), 7.44-7.35 (m, 1H),7.30-7.18 (m, 3H), 5.88 (s, 1H), 4.32 (d, J=5.9 Hz, 2H). ¹³C NMR (101MHz, DMSO-d₆) δ 182.18, 181.88, 162.23 (d, J=244.7 Hz), 148.39, 135.29,133.29, 132.84, 131.28 (d, J=8.9 Hz), 130.83, 128.28 (d, J=2.9 Hz),126.37, 125.86, 124.33 (d, J=9.6 Hz), 118.44 (d, J=22.8 Hz), 116.52 (d,J=21.1 Hz), 101.84, 86.50, 82.29 (d, J=3.3 Hz), 32.34.

2-((3-(3-Methoxyphenyl)prop-2-yn-1-yl)amino)naphthalene-1,4-dione (10)Yield: 92.4 mg (46.2%) of bright orange powder. ESI-MS m/z: 318.25[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.04-7.89 (m, 3H), 7.84 (td, J=7.5,1.4 Hz, 1H), 7.75 (td, J=7.5, 1.3 Hz, 1H), 7.32-7.22 (m, 1H), 7.03-6.92(m, 3H), 5.88 (s, 1H), 4.31 (d, J=6.0 Hz, 2H), 3.74 (s, 3H). ¹³C NMR(101 MHz, DMSO-d₆) δ 182.17, 181.92, 159.55, 148.43, 135.33, 133.31,132.87, 130.85, 130.32, 126.39, 125.87, 124.26, 123.45, 116.65, 115.60,101.80, 85.15, 83.48, 55.63, 32.38.

2-((3-(Pyridin-3-yl)prop-2-yn-1-yl)amino)naphthalene-1,4-dione (11)Yield: 86 mg (46.9%) of yellow powder. ESI-MS m/z: 289.25 [M+H]⁺. ¹H NMR(400 MHz, DMSO-d₆) δ 8.65-8.59 (m, 1H), 8.55 (dd, J=4.9, 1.7 Hz, 1H),8.03-7.89 (m, 3H), 7.88-7.79 (m, 2H), 7.74 (td, J=7.5, 1.3 Hz, 1H), 7.40(ddd, J=8.0, 4.9, 0.9 Hz, 1H), 5.89 (s, 1H), 4.35 (d, J=5.9 Hz, 2H). ¹³CNMR (101 MHz, DMSO-d₆) δ 182.20, 181.89, 152.12, 149.48, 148.42, 139.16,135.31, 133.30, 132.87, 130.85, 126.39, 125.87, 124.04, 119.48, 101.87,88.66, 80.46, 32.41.

2-((3-(4-(Trifluoromethyl)phenyl)prop-2-yn-1-yl)amino)naphthalene-1,4-dione(12) This synthesis was carried out on a 100 mg (0.422 mmol) scale.Yield: 19.8 mg (13.2%) of yellowish-brown powder. ESI-MS m/z: 356.17[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.04-7.90 (m, 3H), 7.85 (td, J=7.5,1.4 Hz, 1H), 7.80-7.70 (m, 3H), 7.64 (d, J=8.1 Hz, 2H), 5.90 (s, 1H),4.36 (d, J=6.0 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 182.21, 181.90,148.44, 137.34-128.87 (m), 128.40-123.88 (m), 101.89, 88.28, 82.21,32.39.

2-((3-(3-(Trifluoromethyl)phenyl)prop-2-yn-1-yl)amino)naphthalene-1,4-dione(13) Yield: 63.3 mg (28.2%) of greenish-yellow powder. ESI-MS m/z:356.17 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.00 (dd, J=7.6, 1.3 Hz, 1H),7.98-7.92 (m, 2H), 7.84 (td, J=7.5, 1.4 Hz, 1H), 7.78-7.70 (m, 4H),7.65-7.57 (m, 1H), 5.89 (s, 1H), 4.34 (d, J=5.9 Hz, 2H). ¹³C NMR (101MHz, DMSO-d₆) δ 182.21, 181.89, 148.43, 137.84-121.51 (m), 101.85,87.25, 81.97, 32.36.

N-(3-(3-((1,4-dioxo-1,4-dihydronaphthalen-2-yl)amino)prop-1-yn-1-yl)phenyl)methanesulfonamide(14) This synthesis was carried out on a 125 mg (0.527 mmol) scale.Yield: 39.3 mg (19.6%) of yellowish powder. ESI-MS m/z: 381.25 [M+H]⁺.¹H NMR (400 MHz, DMSO-d₆) δ 9.87 (s, 1H), 8.00 (dd, J=7.7, 1.3 Hz, 1H),7.96 (dd, J=7.7, 1.3 Hz, 1H), 7.92 (t, J=6.0 Hz, 1H), 7.84 (td, J=7.5,1.4 Hz, 1H), 7.75 (td, J=7.5, 1.4 Hz, 1H), 7.38-7.28 (m, 1H), 7.25-7.20(m, 2H), 7.15 (dt, J=7.6, 1.3 Hz, 1H), 5.88 (s, 1H), 4.32 (d, J=6.0 Hz,2H), 2.99 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 182.17, 181.91, 148.46,139.17, 135.33, 133.32, 132.88, 130.87, 130.27, 127.20, 126.41, 125.87,123.28, 122.36, 120.28, 101.79, 85.68, 83.04, 39.82, 32.36.

2-(Pent-3-yn-1-ylamino)naphthalene-1,4-dione (15) This synthesis wascarried out on a 281 mg (1.20 mmol) scale. Yield: 112 mg (39.1%) ofyellow powder. ESI-MS m/z: 240.17 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ7.98 (dd, J=7.6, 1.3 Hz, 1H), 7.94 (dd, J=7.6, 1.3 Hz, 1H), 7.83 (td,J=7.6, 1.3 Hz, 1H), 7.72 (td, J=7.5, 1.4 Hz, 1H), 7.50 (t, J=6.1 Hz,1H), 5.72 (s, 1H), 3.30 (q, J=6.8 Hz, 2H), 2.45 (tq, J=7.3, 2.5 Hz, 2H),1.74 (t, J=2.6 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 181.92, 181.85,148.66, 135.32, 133.49, 132.67, 130.77, 126.35, 125.79, 100.25, 77.72,77.12, 41.64, 18.07, 3.63.

2-(Hex-3-yn-1-ylamino)naphthalene-1,4-dione (16) This synthesis wascarried out on a 303 mg (1.28 mmol) scale. Yield: 51.2 mg (24.0%) ofyellow powder. ESI-MS m/z: 254.17 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ7.98 (dd, J=7.6, 1.3 Hz, 1H), 7.94 (dd, J=7.7, 1.3 Hz, 1H), 7.83 (td,J=7.5, 1.4 Hz, 1H), 7.73 (td, J=7.5, 1.4 Hz, 1H), 7.50 (t, J=6.2 Hz,1H), 5.74 (s, 1H), 3.30 (q, J=6.9 Hz, 2H), 2.46 (tt, J=7.1, 2.4 Hz, 2H),2.12 (qt, J=7.8, 2.7 Hz, 2H), 1.02 (t, J=7.5 Hz, 3H). ¹³C NMR (101 MHz,DMSO-d₆) δ 181.95, 181.84, 148.71, 135.34, 133.50, 132.68, 130.77,126.36, 125.79, 100.31, 83.58, 77.33, 41.59, 18.21, 14.44, 12.23.

2-(Isopentylamino)naphthalene-1,4-dione (17) (Wang et al., 2014;Gornostaev et al., 2016; Novel tetracyclonaphthooxazole derivative andpreparation method thereof, 2015) Yield: 26 mg (16.9%) of deeporange-red solid. ESI-MS m/z: 244.25 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ7.97 (dd, J=7.7, 1.3 Hz, 1H), 7.94 (dd, J=7.7, 1.3 Hz, 1H), 7.82 (td,J=7.6, 1.4 Hz, 1H), 7.72 (td, J=7.5, 1.3 Hz, 1H), 7.56 (t, J=6.1 Hz,1H), 5.65 (s, 1H), 3.18 (q, J=6.9 Hz, 2H), 1.63 (dp, J=13.2, 6.6 Hz,1H), 1.47 (q, J=7.0 Hz, 2H), 0.91 (d, J=6.6 Hz, 6H). ¹³C NMR (101 MHz,DMSO-d₆) δ 182.03, 181.60, 148.90, 135.28, 133.67, 132.55, 130.85,126.33, 125.77, 99.56, 40.66, 36.44, 25.97, 22.82.

Rac-2-((1-hydroxyhex-4-yn-2-yl)amino)naphthalene-1,4-dione (18) Thissynthesis was carried out on a 200 mg (0.844 mmol) scale. Yield: 66.8 mg(29.4%) of orange powder. ESI-MS m/z: 270.25 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 8.00 (dd, J=7.7, 1.3 Hz, 1H), 7.95 (dd, J=7.7, 1.3 Hz, 1H),7.84 (td, J=7.5, 1.4 Hz, 1H), 7.74 (td, J=7.5, 1.4 Hz, 1H), 6.93 (d,J=8.0 Hz, 1H), 5.80 (s, 1H), 5.00 (t, J=5.5 Hz, 1H), 3.67-3.47 (m, 2H),2.49-2.43 (m, 2H), 1.73 (t, J=2.5 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ182.03, 181.94, 148.40, 135.43, 133.41, 132.77, 130.72, 126.43, 125.81,100.71, 77.96, 76.41, 61.72, 53.82, 20.36, 3.64.

Rac-2-(but-3-yn-2-ylamino)naphthalene-1,4-dione (19) This synthesis wascarried out on a 200 mg (0.844 mmol) scale. Yield: 74.8 mg (39.4%) ofyellow powder. ESI-MS m/z: 226.17 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ7.99 (dd, J=7.8, 1.3 Hz, 1H), 7.94 (dd, J=7.6, 1.3 Hz, 1H), 7.84 (tt,J=7.6, 1.4 Hz, 1H), 7.74 (td, J=7.5, 1.4 Hz, 1H), 7.50 (d, J=7.3 Hz,1H), 5.84 (d, J=0.6 Hz, 1H), 4.41 (pd, J=7.1, 2.2 Hz, 1H), 3.34 (d,J=2.2 Hz, 1H), 1.51 (d, J=6.9 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ182.24, 181.87, 147.63, 135.31, 134.92, 134.03, 133.13, 132.91, 130.81,126.53, 126.40, 125.96, 125.82, 110.63, 102.45, 99.98, 83.65, 74.27,20.71, 14.31.

Rac-2-(pent-4-yn-2-ylamino)naphthalene-1,4-dione (20) This synthesis wascarried out on a 200 mg (0.844 mmol) scale. Yield: 26 mg (12.9%) ofyellow powder. ESI-MS m/z: 240.25 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ7.98 (dd, J=7.7, 1.3 Hz, 1H), 7.94 (dd, J=7.6, 1.3 Hz, 1H), 7.83 (td,J=7.5, 1.4 Hz, 1H), 7.72 (td, J=7.5, 1.4 Hz, 1H), 7.15 (d, J=8.9 Hz,1H), 5.76 (d, J=0.7 Hz, 1H), 3.76 (dq, J=8.9, 6.4 Hz, 1H), 2.91 (t,J=2.7 Hz, 1H), 2.61-2.41 (m, 2H), 1.26 (d, J=6.4 Hz, 3H). ¹³C NMR (101MHz, DMSO-d₆) δ 181.99, 181.96, 147.86, 135.35, 133.41, 132.70, 130.75,126.38, 125.78, 100.57, 81.76, 73.44, 47.14, 24.73, 19.18.

Rac-2-((1-phenylbut-3-yn-1-yl)amino)naphthalene-1,4-dione (21) Yield: 45mg (23.6%) of brownish yellow powder. ESI-MS m/z: 302.25 [M+H]⁺. ¹H NMR(400 MHz, DMSO-d₆) δ 7.99 (dd, J=7.7, 1.3 Hz, 1H), 7.88 (dd, J=7.7, 1.4Hz, 1H), 7.84 (d, J=7.9 Hz, 1H), 7.80 (td, J=7.5, 1.4 Hz, 1H), 7.72 (td,J=7.5, 1.4 Hz, 1H), 7.53-7.44 (m, 2H), 7.40-7.30 (m, 2H), 7.34-7.22 (m,1H), 5.60 (s, 1H), 4.72 (td, J=8.0, 6.0 Hz, 1H), 2.99 (ddd, J=16.7, 8.1,2.6 Hz, 1H), 2.94-2.90 (m, 1H), 2.78 (ddd, J=16.7, 6.0, 2.7 Hz, 1H),2.08 (s, 1H). ¹³C NMR (101 MHz, DMSO-d₆) δ 182.02, 181.90, 147.98,140.90, 135.34, 133.17, 132.83, 130.71, 128.91, 128.09, 127.24, 126.39,125.79, 101.89, 81.63, 73.65, 55.56, 26.49, 1.64.

(E)-2-((3,7-Dimethylocta-2,6-dien-1-yl)amino)naphthalene-1,4-dione (22)This synthesis was carried out on a 200 mg (0.844 mmol) scale. Yield:78.1 mg (29.9%) of bright orange solid. ESI-MS m/z: 310.17 [M+H]⁺. ¹HNMR (400 MHz, DMSO-d₆) δ 7.97 (dd, J=7.7, 1.3 Hz, 1H), 7.93 (dd, J=7.7,1.2 Hz, 1H), 7.82 (td, J=7.5, 1.4 Hz, 1H), 7.72 (td, J=7.5, 1.4 Hz, 1H),7.65 (t, J=6.0 Hz, 1H), 5.57 (s, 1H), 5.19 (tq, J=6.3, 1.3 Hz, 1H), 5.04(ddp, J=7.0, 5.7, 1.5 Hz, 1H), 3.80 (t, J=6.2 Hz, 2H), 2.14-1.93 (m,4H), 1.70 (d, J=1.3 Hz, 3H), 1.58 (d, J=1.4 Hz, 3H), 1.54 (d, J=1.3 Hz,3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 182.08, 181.59, 148.75, 138.68,135.27, 133.62, 132.58, 131.38, 130.82, 126.30, 125.79, 124.23, 120.09,100.19, 40.47, 26.24, 25.88, 18.01, 16.65.

(E)-2-Bromo-3-((3,7-dimethylocta-2,6-dien-1-yl)amino)naphthalene-1,4-dione(23) This compound was isolated as a byproduct during purification of 17and purified by standard phase chromatography on a pre-packed silica gelcolumn eluting with a hexanes:ethyl acetate gradient. Yield: 53.1 mg(16.3%) of deep red powder. ESI-MS m/z: 388.00 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 8.05-7.93 (m, 2H), 7.82 (td, J=7.5, 1.5 Hz, 1H), 7.74 (td,J=7.5, 1.4 Hz, 1H), 7.44-7.35 (m, 1H), 5.36-5.25 (m, 1H), 5.08-4.97 (m,1H), 4.36 (t, J=6.2 Hz, 2H), 2.14-1.92 (m, 4H), 1.68 (d, J=1.3 Hz, 3H),1.56 (s, 3H), 1.52 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 180.33, 138.03,135.27, 133.08, 131.32, 127.01, 126.49, 124.25, 122.45, 26.25, 25.86,18.01, 16.74.

Synthesis of(E)-2-((3,7-Dimethylocta-2,6-dien-1-yl)amino)-3-methylnaphthalene-1,4-dione(24) To 2-methyl-1,4-naphthoquinone (302 mg, 1.6 mmol) inmethanol:dichloromethane (2 mL each) was added geranylamine (396 mg,2.58 mmol) and the mixture was stirred at room temperature for 48 h. Thesolution was concentrated in vacuo and the reaction was purified on C18silica gel eluting with a water:acetonitrile gradient to yield 55 mg ofa deep, red oil (11% yield). ESI-MS m/z: 324.17 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 7.92 (ddd, J=7.5, 4.4, 1.3 Hz, 2H), 7.77 (td, J=7.5, 1.4 Hz,1H), 7.68 (td, J=7.5, 1.3 Hz, 1H), 6.68 (t, J=6.6 Hz, 1H), 5.32-5.20 (m,1H), 5.02 (tdd, J=6.8, 3.0, 1.5 Hz, 1H), 4.14 (t, J=6.4 Hz, 2H), 2.10(s, 3H), 2.07-1.88 (m, 4H), 1.65 (s, 3H), 1.57 (s, 3H), 1.52 (s, 3H).¹³C NMR (101 MHz, DMSO-d₆) δ 182.65, 182.14, 147.10, 137.47, 134.86,133.21, 132.55, 131.33, 130.70, 126.03, 125.82, 124.25, 123.46, 111.43,42.85, 39.25, 26.21, 25.86, 18.00, 16.62, 10.86.

iii. Synthesis of 2-alkoxy-1,4-naphthoquinones, General Procedure

2-hydroxy-1,4-naphthoquinone was dissolved in DMF in a round bottomflask charged with a stir bar. To this was added the appropriatetosylate reagent as a solution in DMF. With stirring, solid K₂CO₃ wasadded to the flask; at this point, the color of the reaction mixturechanged from pale yellow to deep red in color. The reaction was thenrefluxed under an argon atmosphere for 2 hr, at which time the reactionwas cooled, poured into a separatory funnel with DI H₂O and DCM, andextracted. The organic layer was washed 2×20 mL with concentratedNaHCO₃, 2×15 mL brine, then dried over anhydrous MgSO₄. The solid dryingagent was removed by gravity filtration and solvent removed by rotaryevaporation to yield thick, red oil, which was then purified byreversed-phase chromatography.

2-(Isopentyloxy)naphthalene-1,4-dione (25) (Adin and Fleming, 1980) Thissynthesis was carried out on a 160 mg (0.920 mmol) scale. Yield: 21.2 mg(9.4%) of off-white powder. ESI-MS m/z: 245.08 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 8.04-7.93 (m, 2H), 7.91-7.75 (m, 2H), 6.38 (s, 1H), 4.08 (t,J=6.6 Hz, 2H), 1.78 (hept, J=13.2, 6.6 Hz, 1H), 1.67 (q, J=6.7 Hz, 2H),0.94 (d, J=6.6 Hz, 6H). ¹³C NMR (101 MHz, DMSO-d₆) δ 184.98, 180.04,160.11, 134.91, 134.01, 131.99, 131.31, 126.51, 125.96, 110.68, 68.32,37.00, 25.07, 22.79.

Alternate synthesis of 2-(Isopentyloxy)naphthalene-1,4-dione (25) Amodified procedure adapted from literature syntheses of2-(methoxy)naphthalene-1,4-dione was also used to synthesize 25 inlarger amounts (Ogata et al., 2016; Sreelatha et al., 2014).2-hydroxy-1,4-naphthoquinone (348 mg, 2 mmol) was added to isoamylalcohol (7 mL) and stirred. 0.3 mL of concentrated HCl was then addedand the reaction mixture was refluxed for 4 hr. The deep red mixture wascooled to room temperature and then further in a refrigerator overnight.The solid precipitate was isolated by filtration, dissolved in DCM, andwash with K₂CO₃ (3×30 mL) to removed 3-substituted-2-hydroxy sideproducts. The pooled aqueous layers were back extracted 2×30 mL withDCM, the organic layers pooled and washed with brine, then dried overMgSO₄. Following filtration and removal of solvent in vacuo, the dark,oily crude product was purified by reversed-phase chromatography usingpre-packed C18 silica columns. Yield: 157 mg (32.1%) of tan solid withthe same analytical data as above.

2-Butoxynaphthalene-1,4-dione (26) (Fieser, 1926; Wang et al., 2015)This synthesis was carried out on a 169 mg (0.971 mmol) scale. Yield: 27mg (12.1%) of yellow powder. ESI-MS m/z: 231.17 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 7.99 (dd, J=6.9, 2.0 Hz, 1H), 7.97-7.93 (m, 1H), 7.89-7.79(m, 2H), 6.34 (s, 1H), 4.04 (t, J=6.5 Hz, 2H), 1.75 (p, J=8.4, 6.5 Hz,2H), 1.44 (h, 2H), 0.94 (t, J=7.4 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ184.94, 180.03, 160.09, 134.90, 134.01, 131.96, 131.28, 126.51, 125.95,110.61, 69.44, 30.35, 19.08, 14.04.

2-(Benzyloxy)naphthalene-1,4-dione (27) (Kumar et al., 2017; Ogata etal., 2016) This synthesis was carried out on a 169 mg (0.971 mmol)scale. Yield: 32.6 mg (12.7%) of off-white powder. ESI-MS m/z: 265.17[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 8.05-7.94 (m, 2H), 7.91-7.78 (m,2H), 7.53-7.45 (m, 2H), 7.50-7.35 (m, 3H), 6.49 (s, 1H), 5.18 (s, 2H).¹³C NMR (101 MHz, DMSO-d₆) δ 184.98, 180.01, 159.73, 135.45, 134.97,134.12, 131.92, 131.30, 129.07, 128.96, 128.73, 126.57, 126.02, 111.28,71.19.

2-(But-2-yn-1-yloxy)naphthalene-1,4-dione (28) This synthesis wascarried out on a 160 mg (0.920 mmol) scale. Yield: 39.3 mg (19.2%) ofoff-white powder. ESI-MS m/z: 223.08 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ8.05-7.91 (m, 2H), 7.91-7.79 (m, 2H), 6.39 (s, 1H), 4.88 (q, J=2.4 Hz,2H), 1.90 (t, J=2.4 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 184.84,179.80, 158.79, 135.00, 134.17, 131.84, 131.23, 126.58, 126.02, 111.51,86.09, 73.11, 58.05, 3.66.

2-(Pent-2-yn-1-yloxy)naphthalene-1,4-dione (29) This synthesis wascarried out on a 169 mg (0.971 mmol) scale. Yield: 64 mg (27.5%) ofoff-white powder. ESI-MS m/z: 241.17 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ8.05-7.91 (m, 2H), 7.92-7.79 (m, 2H), 6.39 (s, 1H), 4.89 (t, J=2.2 Hz,2H), 2.28 (qt, J=7.5, 2.2 Hz, 2H), 1.09 (t, J=7.5 Hz, 3H). ¹³C NMR (101MHz, DMSO-d₆) δ 184.84, 179.79, 158.78, 135.01, 134.18, 131.82, 131.23,126.59, 126.02, 111.52, 91.46, 73.33, 58.07, 13.86, 12.19.

2-(Hex-2-yn-1-yloxy)naphthalene-1,4-dione (30) This synthesis wascarried out on a 160 mg (0.920 mmol) scale. Yield: 21.1 mg (9.0%) ofoff-white powder. ESI-MS m/z: 255.25 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ8.03-7.99 (m, 1H), 7.99-7.95 (m, 1H), 7.91-7.80 (m, 2H), 6.40 (s, 1H),4.91 (t, J=2.2 Hz, 2H), 2.26 (tt, J=7.0, 2.2 Hz, 2H), 1.48 (h, J=7.2 Hz,2H), 0.93 (t, J=7.3 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 184.82,179.82, 158.75, 135.02, 134.19, 131.81, 131.23, 126.59, 126.03, 111.61,90.05, 74.13, 58.05, 21.76, 20.38, 13.68.

2-(Hept-2-yn-1-yloxy)naphthalene-1,4-dione (31) This synthesis wascarried out on a 169 mg (0.971 mmol) scale. Yield: 15.9 mg (6.1%) ofoff-white powder. ESI-MS m/z: 269.25 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ8.06-7.92 (m, 2H), 7.86 (pd, J=7.3, 1.7 Hz, 2H), 6.40 (s, 1H), 4.91 (t,J=2.2 Hz, 2H), 2.28 (tt, J=7.0, 2.2 Hz, 2H), 1.52-1.27 (m, 4H), 0.85 (t,J=7.2 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 184.82, 179.84, 158.74,135.03, 134.20, 131.82, 131.24, 126.60, 126.03, 111.65, 74.00, 58.05,30.31, 21.74, 18.10, 13.83.

2-((3-Phenylprop-2-yn-1-yl)oxy)naphthalene-1,4-dione (32) This synthesiswas carried out on a 169 mg (0.971 mmol) scale. Yield: 92 mg (32.9%) ofcream-colored solid. ESI-MS m/z: 289.24 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 8.07-8.01 (m, 1H), 8.01-7.96 (m, 1H), 7.92-7.80 (m, 2H),7.55-7.45 (m, 2H), 7.50-7.36 (m, 3H), 6.51 (s, 1H), 5.20 (s, 2H). ¹³CNMR (101 MHz, DMSO-d₆) δ 184.88, 179.77, 158.74, 135.02, 134.21, 132.07,131.86, 131.28, 129.86, 129.28, 126.62, 126.06, 121.56, 111.69, 88.38,83.20, 58.09.

B. Cell Culture

The HT22 neuronal cell line is a subclone of HT4, derived from the mousehippocampus (Morimoto and Koshland, 1990). They do not express activeionotropic glutamate receptors and are not subject to excitotoxicity(Maher and Davis, 1996). The HT22 cells used in this study were kindlyprovided by Dr. David Schubert (The Salk Institute for BiologicalStudies, La Jolla, CA, USA). The cells were grown in Dulbecco's ModifiedEagle's Medium (DMEM/high glucose) supplemented with 10% fetal bovineserum (Hyclone) and 5 mL of antibiotic-antimycotic (Amphotericin B,Penicillin, and Streptomycin; Invitrogen) at 37° C. in 5% CO₂.

C. Cell Viability Assay

HT-22 cells were seeded onto 96-well plates at 2.5×10³ cells per well in75 μL of medium and maintained at 37° C. in 5% CO₂ overnight prior tothe initiation of experimental treatments. For glutamate toxicitytesting, cells were subsequently treated with 25 μL of medium containingglutamate (monosodium glutamate, Sigma, 1 M stock concentration inmedia, to achieve a final concentration of 10 mM) plus compound (stockin DMSO) and maintained at 37° C. in 5% CO₂ for 24 hr. CellTiter-Blue®(resazurin cell viability assay reagent) was then added to each well ata final concentration of 0.125 mg/mL. The mixture was allowed toincubate for 2-4 hr until sufficient color change occurred. Cellviability was measured as a function of resorufin fluorescence intensityusing a Tecan M200 Pro spectrophotometer, 560 nm/590 nm(excitation/emission). Cell viability was calculated as a percentagecompared with untreated controls. EC50s were determined using GraphPadPrism's “log(inhibitor) vs. normalized response-Variable slope”function. Morphology of HT22 cells following treatments was determinedby phase-contrast microscopy.

D. Zebrafish Breeding and Maintenance

Zebrafish (AB strain) were obtained from the Zebrafish InternationalResource Center (supported by P40 RR012546 from NIH-NCRR). Zebrafishwere maintained and crossed according to standard methods (Westerfield,2000). Fertilized eggs were collected and placed in E3 embryo medium andpositioned in an incubator set at 28.5° C. with a 14/10-hr light/darkcycle (Kimmel et al., 1995). To determine the lethal dose of eachcompound, 96-well plates containing one zebrafish (7 dayspost-fertilization, dpf) per well in 100 μL of tank water were used. 100μL of each compound (0.5-15 μM) was added to each well for 12 animals(one row) for a final volume of 200 μL. One row of larvae was used asdimethyl sulfoxide (DMSO)-only controls. The 96-well plate was placed ona warmer at 28.5° C. and fish were observed for changes in phenotype,behavior and mortality initially after addition of compound, after 1 hrtreatment and after 5 hr treatment. All zebrafish studies were approvedby the Medical University of South Carolina Institutional Animal Careand Use Committee (#180278) and performed in accordance with theguidelines.

E. Toxicity Studies in Zebrafish

Using a 96-well plate, 7 dpf zebrafish were placed one per well with 100μL of tank water. 100 μL of each compound was then added to each wellfor 12 animals (one row) for a final volume of 200 μL. One row ofzebrafish larvae was used as DMSO only controls. The 96-well plate wasthen placed on a warmer plate at 28.5° C. and the fish were observed forchanges in phenotype, behavior and mortality initially after addition ofcompound, after 1 hr and 5 hr of treatment.

F. Induction and Monitoring of Seizures in Zebrafish

We induced seizures in 7 dpf zebrafish larvae by the addition of 15 mMPTZ as originally developed by Baraban et al., 2005. In a 48-well plate,one 7-dpf zebrafish was added per well. Larvae were dosed with eachcompound at a sub-lethal dose 1 hr prior to PTZ treatment. Two controlrows were included with each experiment—tank water only control and PTZonly. Seizures were induced by adding PTZ to wells to yield a finalconcentration of 10 mM. After 5 min, the plate was transferred to theDaniovision instrument (Noldus Information Technology) and the chamberlight was turned on. After 2 min, MediaRecorder (Noldus) was used torecord video for 15 min. A small number of videos were acquired at 25frames per second, but the majority of data were acquired at 60 framesper second. After recording, fish were monitored visually for survival,and agitation of the plates to elicit a startle response in zebrafishwould determine whether reduced swim distances were accompanied bynormal behavior and not the result of sedative effects. Ethovision XTsoftware (Noldus) was used to track the fish movement from the videoimages in order to calculate the total distance traveled over 15 min.All experimental comparisons were made between animals from the sameclutch.

G. Stability of Compound 1 in CD-1 Mouse Plasma

The plasma stability of 1 was determined by WuXi AppTec. Three sampleswere prepared using 3 μL working solution (2 μg/mL in acetonitrile:H₂O70:30) spiked in 57 μL CD-1 mouse plasma (EDTA-K₂) and mixed well.Samples were stored at room temperature for two hours, mixed well, thenprecipitated. Three 0 hr samples were prepared in the exact same mannerbut were precipitated immediately upon preparation. Precipitate mixtureswere centrifuged at 4000 rpm for 15 min, then 2 μL supernatant was usedfor LC-MS/MS analysis.

H. In Vivo Pharmacokinetics Procedure in Mice

The PK studies of 1 were performed by WuXi AppTec and PK studies of 3and 25 were performed by Touchstone Biosciences (Peng et al., 2006b;Peng et al., 2001; Peng et al., 2006a; Peng et al., 2009). Male CD-1mice were fed a standard laboratory rodent diet and housed in individualcages on a 12 hr light and 12 hr dark cycle with room temperaturemaintained at 22±3° C. and relative humidity at 50±20%. Animals weretypically fasted overnight before dosing, with food returned after the 6hr blood samples were obtained. Water was provided ad libitum throughoutthe study. The dosing solution of each test compound was prepared in adesired oral or intravenous formulation. Three to four animals weredosed via gavage needle for oral administration at 10-20 mg/kg (10-20mL/kg) or via tail vein injection for IV administration at 2-5 mg/kg(2-5 mL/kg). All blood samples (30-200 μL per sample) were taken viaappropriate vein (saphenous, jugular, or submandibular vein) at 5, 15,and 30 min and 1, 2, 4, 6, 8, and 24 hr after dosing. Fluid replacement(1.5 mL of 0.9% NaCl injection, USP) was administered subcutaneouslyonce after the 2 hr blood sampling. Blood samples were collected in BDMicrotainer tubes coated with anticoagulant, placed on ice, and within30 min, centrifuged at 15,000 g for 5 min to obtain plasma samples. Allplasma samples were stored at −70° C. until analysis.

I. Bioanalysis of Samples.

Plasma samples were prepared as follows. Three volumes of acetonitrilecontaining internal standard was added to one volume of plasma toprecipitate proteins. Samples were centrifuged (3000 g for 10 min) andsupernatant removed for analysis by LC-MS/MS. Calibration standards andquality controls were made by preparation of a 1 mg/mL stock solutionand subsequently a series of working solutions in methanol:water (1:1,v/v) which were spiked into blank plasma to yield a series ofcalibration standard samples in the range of 1.0 ng/mL to 10 μg/mL andquality control samples at three concentration levels (low, middle andhigh). All incurred PK/PD plasma samples were treated identically to thecalibration standards and quality control samples. LC-MS-MS analysis wasperformed utilizing multiple reaction monitoring for detection ofcharacteristic ions for each drug candidate, additional related analytesand internal standard.

J. PK Data Analysis.

Plasma concentrations were measured as described above to determine aconcentration vs. time profile. The area under the plasma concentrationvs time curve (AUC) was calculated using the linear trapezoidal method.Fitting of the data to obtain pharmacokinetic parameters was carried outusing non-compartmental analysis. Key PK parameters reported followingintravenous administration are as follows: terminal half-life t_(1/2),initial plasma concentration C₀, area under the plasma concentration vs.time curve AUC, volume of distribution at steady-state V_(ss), totalplasma clearance C_(Lp), and mean residence time MRT. Key PK parametersreported following extravascular administration are as follows: terminalhalf-life t_(1/2), maximum plasma concentration C_(max), time to reachmaximum plasma concentration t_(max), area under the plasmaconcentration vs. time curve AUC, mean residence time MRT, andbioavailability F. All parameters are expressed for individual animalsas well as mean, standard deviation, and coefficient of variation.

K. Statistical Analyses

Statistical analyses were performed with GraphPad Prism 6 software. Forthe in vitro HT22 oxytosis assay, dose-response data were fit usingGraphPad Prism's log(inhibitor) vs normalized response-variable slopeparameters to determine EC50 values and 95% confidence intervals. Forthe zebrafish locomotive anti-seizure activity data, multiplecomparisons were made using a one-way analysis of variance (ANOVA) witha Kruskal-Wallis Test, followed by the Dunn's Method to determinesignificant differences between all pairs or between control andexperimental groups using the Dunn Method for Joint Ranking. Differenceswere considered statistically significant when p<0.05. Data fromzebrafish experiments are represented as scatter dot plots of individualmeasurements, with means±standard deviation indicated by bars.

The following compounds may be synthesized based on the syntheticschemes described above.

Example 3

Treatment of Medication-Resistant Epilepsy

From preliminary testing of four previous compounds of interest in the 6Hz mouse seizure model, two compounds of particular interest, compound 3(NT-181) and compound 17 (NT-102), displayed protection against seizureswith few or no toxic events, as shown by Rotarod assay and behavioralobservations. Synthesis for both compounds has been successfully scaledup. The pharmacokinetic (PK) parameters of these two Vitamin K analogswere determined via intravenous, intraperitoneal and oral administrationin mice, and preliminary in vitro ADMET parameters were assessed (Table3). These compounds are Ames-negative, have hepatic stability greaterthan 1 hour, can be formulated in oral solution and IV injectionsolution, can be synthesized in greater than 1 mg quantities, and haveestablished brain PK. However, brain tissue binding was high for bothcompounds (97-98%). Both 3 (NT-181) and 17 (NT-102) have been tested forefficacy in the 6 Hz mouse seizure model at 22 mA and 32 mA, the cornealkindled mouse model, and the rat in vitro spontaneous electrographicbursting model of pharmacoresistance (Table 4). During the course ofthis testing, it was found that there were solubility issues with theformulation used (5% DMSO:95% Neobee); sacrifice of the animals revealedvisible intraperitoneal deposits of the compound in the cavity aroundthe injection site, and thus some of the compound may not be gettingpast this site. Higher concentrations may be required to complete thetitration curves for protection in the 6 Hz mouse seizure model at 32 mAand 44 mA. Going through the records of the first time the Vitamin Kanalogs were tested in the 6 Hz model (2013-2014) revealed that theNINDS ETSP/University of Utah Anticonvulsant Drug Development (ADD)program had used a type of Miglyol for formulation. However, the type ofMiglyol was not noted. Solubility of the Vitamin K analogs were testedwith different types of Miglyol with varying viscosities and it wasfound that Miglyol840 (the least viscous Miglyol) was best. Preliminarystudies with this new formulation reveal excellent solubility with fullprotection in the 6 Hz mouse seizure model at 22 mA, and quantificationof both compounds (using this new formulation) in this model at 22 mA,32 mA and 44 mA, is being performed as well as retesting in the cornealkindled model. Protection in acute and chronic seizure mouse models fromthe ADD workflow for pharmacoresistant epilepsy will be tested. Rat PKassessments will be performed, and seizure protection assessed in ratmodels in the ADD workflow such as the 6 Hz model andlamotrigine-resistant amygdala kindled mode. Results are shown below inTable 3 and Table 4.

TABLE 3 Mouse PK and in vitro ADMET summary NT-181 NT-102 StandardStandard Parameter Mean deviation Mean deviation IntravenousAdministration  5 mg/kg  5 mg/kg (20% DMA: 40% PEG300: 40% H₂O) T_(1/2)(hr) 14.7 3.78 4.47 0.536 C₀ (ng/mL) 1523 254 1907 84.7 C_(last) (ng/mL)1.37 0.167 0.154 0.0283 AUC_(last) (hr*ng/ml) 421 64.8 636 49.3AUC_(inf) (hr*ng/mL) 448 69 637 49.1 MRT_(inf) (hr) 4.28 1.14 0.8740.0365 AUC_(inf)/D (hr*mg/mL) 89.6 13.7 127 9.82 CL_(p) (ML/min/kg) 18827.3 131 10 V_(ss) (L/kg) 48.6 15.9 6.89 0.512 Brain Plasma Ratio 0.3730.816 Intraperitoneal Admin. 20 mg/kg 20 mg/kg (5% DMSO: 95% neobee)T_(1/2) (hr) 7.25 0.814 3.38 0.162 T_(max) (hr) 1 0 1 0 C_(max) (ng/mL)147 33.5 320 91.1 C_(max)/D (kg/kL) 7.33 1.67 16 4.56 t_(last) (hr) 24 024 0 C_(last) (ng/ml) 1.47 0.312 1.95 0.276 AUC_(last) (hr*ng/ml) 46328.6 1716 265 AUC_(inf) (hr*ng/mL) 478 28 1726 265 AUC_(inf)/D(hr*mg/kL) 23.9 1.4 86.3 13.2 MRT_(inf) (hr) 4.2 0.696 4.39 0.49 OralAdministration 20 mg/kg 20 mg/kg (5% DMSO: 95% Neobee) t_(1/2) (hr) 3.340.183 5.28 0.552 t_(max) (hr) 4.0 0 0.25 0 C_(max) (ng/mL) 90.2 12.81025 273 C_(max)/D (kg/kL) 4.51 0.639 51.2 13.7 t_(last) (hr) 24 0 24 0C_(last) (ng/mL) 1.21 0.218 1.43 0.278 AUC_(last) (hr*ng/ml) 682 135 903175 AUC_(inf) (hr*kg/mL) 688 135 914 177 AUC_(inf)/D (hr*kg/kL) 34.46.76 45.7 8.83 MRT_(inf) (hr) 5.71 0.29 2.50 0.228 F (%) 38.4 7.54 35.96.93 In vitro ADMET Brain tissue binding 97.4 0.486 98.5 0.168 (% bound)Aqueous solubility at 4.79 9.64 pH 7.4 (ug/mL in 50 mM phosphate buffer)Liver microsome >1 hr >1 hr stability (t_(1/2)) Mini-Ames test NegativeNegative (mutagenicity)

Table 2 provides a summary of efficacy studies in vivo in mice, and inrat brain slices, using the 5% DMSO:95% Neobee formulation. Use of 3(NT-181) showed pigmented urine due to the color of the compound itself.Pigmented urine was not seen with 17 (NT-102). Injection issues with theNeobee oil formulation were observed. The injection was reformulatedwith Miglyol840 oil mix and a lower EC₅₀ was observed.

TABLE 4 Summary of efficacy studies in vivo in mice, and in rat brainslices, using the 5% DMSO: 95% Neobee formulation. Number of AnimalsProtected Model tested NT-181 NT-102 6 Hz mouse seizure model, 22 mA 8/8at 100 mg/kg 8/8 at 400 mg/kg Time of Peak Effect 0.5 hours 0.25 hoursEffective Dose 80.60 (25-100) mg/kg 209.05 (25-400) mg/kg 6 Hz mouseseizure model, 32 mA 0/7 at 100 mg/kg; 3/8 at 300 mg/kg 2/8 at 400 mg/kg6 Hz mouse seizure model, 44 mA 0/8 at 100 mg/kg Neurotoxicity (Rotorodassay) 1/71 showed toxicity 0/72 showed toxicity Mouse corneal kindledmodel 0/8 at 300 mg/kg 0/24 at 100, 300 and 400 mg/kg Rat in vitrospontaneous 0/8 at 5 uM 0/9 at 10 uM electrographic bursting model

Additional rodent studies showed that compound 17 (NT-102) had efficacyin protecting against electrically-induced seizures, and separatestudies were performed to see if NT-102 would affect motor functionusing the rotarod test in the same mouse or rat. Mice or rats wereadministered the NT-102 compound i.p., one hour was allowed to pass(since the time to peak effect was determined to be about one hour), andthen rodents were then placed on the Rotarod to determine if any motordefects would be observed; after the rotarod test, then rodents wereadministered an electrical stimulation to determine if the dose ofNT-102 would protect against (i.e., prevent) the seizures due to theelectrical stimulation. In particular, using (i) electrical model totest for seizures in mice and, separately, (ii) in rat 6 Hz 40V model.More specifically, NT-102 was injected (i.p.) in 5% DMSO/95% Miglyol 840as vehicle. Mice were challenged with 6 Hz 44 mA current to see ifseizures would result. The Effective Dose (ED₅₀) of NT-102 to preventseizures was observed to be 263.74 mg/kg (with a 95% confidence intervalof 198.86-321.76 mg/kg). The Time of Peak Effect (TPE) was observed 1hour. Additional data is shown below in Table 5. The mice wereadministered the NT-102 at various doses and placed on the rotarod(i.e., “Rotarod (Tox)”) to test if the mice would fall off as a model ofdue to the effects of NT-102; for the “Rotarod(Tox)” results, the “N/T”indicates the number of mice that fell off the rotarod (e.g., forRotarod(Tox) results, if N/T is 0/8, then zero out of eight mice felloff). For the electrical stimulation data (i.e., “6 Hz 44 mA”), anelectrical stimulation of 6 Hz 44 mA was administered. For the 6 Hz 44mA data, “N/T” refers to the number of mice who were protected againstthe seizures (e.g., for the 6 Hz 44 mA results if N/T is 4/8, then fourout of eight mice tested were protected against the seizures anddisplayed no seizures due to the electrical current application).Additional electrical dosages and timepoints were tested in mice orrats, as shown in the tables below. The 44 mA electrical dosage was usedsince it is considered to be a model to test for medication-resistantepilepsy.

TABLE 5 Results with NT-102. Test Dose Time N/T Rotarod (Tox) 100 mg/kg1 hr 0/8 6 Hz 44 mA 100 mg/kg 1 hr 0/8 Rotarod (Tox) 200 mg/kg 1 hr 2/86 Hz 44 mA 200 mg/kg 1 hr 2/8 Rotarod (Tox) 300 mg/kg 1 hr 0/8 6 Hz 44mA 300 mg/kg 1 hr 4/8 Rotarod (Tox) 400 mg/kg 1 hr 1/8 6 Hz 44 mA 400mg/kg 1 hr 8/8A different 32 mA electrical stimulation was used in additional studieswith mice, as shown below. Results are shown in Table 6. As expected, alower ED50 dose was observed of 152.72 mg/kg (with a 95% confidenceinterval of 98.63-210.98 mg/kg) was observed using this lower amount ofelectrical stimulation.

TABLE 6 Results with NT-102. Test Dose Time N/T Rotarod (Tox)  50 mg/kg1 hr 0/8 6 Hz 32 mA  50 mg/kg 1 hr 0/8 Rotarod (Tox) 100 mg/kg 1 hr 0/86 Hz 32 mA 100 mg/kg 1 hr 4/8 Rotarod (Tox) 200 mg/kg 1 hr 1/8 6 Hz 32mA 200 mg/kg 1 hr 3/8 Rotarod (Tox) 300 mg/kg 1 hr 3/8 6 Hz 32 mA 300mg/kg 1 hr 7/8Rats were tested using a motor function test (referred to as “MMI(Tox)”to test for minimal motor impairment, as shown in the tables below)using dosages of NT-102 administered i.p., followed by electricalstimulation of 6 Hz 40 V. Time to peak result was observed to be aboutone hour. Results are shown in Table 7, below.

TABLE 7 Results with NT-102. Test Dose Time N/T MMI (Tox) 300 mg/kg 0.25hr 0/4 6 Hz 40 V 300 mg/kg 0.25 hr 3/8 MMI (Tox) 300 mg/kg  0.5 hr 0/4 6Hz 40 V 300 mg/kg  0.5 hr 0/8 MMI (Tox) 300 mg/kg    1 hr 0/4 6 Hz 40 V300 mg/kg    1 hr 5/8 MMI (Tox) 300 mg/kg    2 hr 0/4 6 Hz 40 V 300mg/kg    2 hr 0/4 MMI (Tox) 300 mg/kg    4 hr 0/4 6 Hz 40 V 300 mg/kg   4 hr 0/4Additional studies with the rat model were performed as shown in Table8, below. The ED50 in the rats for the 6 Hz 40 V electrical stimulationwas observed to be 293 mg/kg (with a 95% confidence interval of 242-441mg/kg). The 6 Hz 40 V electrical stimulation amount was used becausethis electrical dosage is considered to be a rat model formedication-resistant epilepsy.

TABLE 8 Results with NT-102. Test Dose Time N/T 6 Hz 40 V 200 mg/kg 1 hr1/8 6 Hz 40 V 250 mg/kg 1 hr 2/8 6 Hz 40 V 350 mg/kg 1 hr 6/8 MMI (Tox)350 mg/kg 1 hr 0/8

Example 4

Treatment of Parkinson's Disease

Pink1 mutant zebrafish were obtained from Dr. Daniel Hesselson from theGarvan Institute, Sydney, Australia. His paper in Cell Chemical Biology(2017) outlines the phenotypic screening strategy; one of these factorstested is touch-evoked escape response—young zebrafish are placed inmulti-well plates and incubated with a mitochondrial toxin, rotenone. WTzebrafish with or with rotenone treatment, as well as untreated pink−/−zebrafish are able to react to a touch to their tail by theinvestigator. However, pink1−/− zebrafish treated with rotenone have asignificantly dampened touch response, which is rescued by pretreatmentwith 1 μM 1st generation VK analog NT-108. While one concentration hasbeen tested, the 2nd-generation VK analogs are being tested on thisanimal model of Parkinson's. Results are shown in FIG. 6 .

All of the methods disclosed and claimed herein can be made and executedwithout undue experimentation in light of the present disclosure. Whilethe compositions and methods of this invention have been described interms of preferred embodiments, it will be apparent to those of skill inthe art that variations may be applied to the methods and in the stepsor in the sequence of steps of the method described herein withoutdeparting from the concept, spirit and scope of the invention. Morespecifically, it will be apparent that certain agents which are bothchemically and physiologically related may be substituted for the agentsdescribed herein while the same or similar results would be achieved.All such similar substitutes and modifications apparent to those skilledin the art are deemed to be within the spirit, scope and concept of theinvention as defined by the appended claims.

REFERENCES

The following references, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference.

-   U.S. Pat. No. 5,399,363-   U.S. Pat. No. 5,466,468-   U.S. Pat. No. 5,543,158-   U.S. Pat. No. 5,580,579-   U.S. Pat. No. 5,629,001-   U.S. Pat. No. 5,641,515-   U.S. Pat. No. 5,725,871-   U.S. Pat. No. 5,756,353-   U.S. Pat. No. 5,780,045-   U.S. Pat. No. 5,792,451-   U.S. Pat. No. 5,804,212-   U.S. Pat. No. 6,613,308-   Adin and Fleming, CO(III) Complex containing radiation sensitive    element with diazo recording layer. Eastman Kodak Co., U.S. Pat. No.    4,195,998A, 1980.-   Albrecht et al., Mechanisms of oxidative glutamate toxicity: the    glutamate/cystine antiporter system xc- as a neuroprotective drug    target. CNS Neurol Disord Drug Targets; 9(3):373-82, 2010.-   Alsdorf and Wyszynski, Teratogenicity of sodium valproate. Expert    Opin Drug Saf.; 4(2):345-53, 2005.-   Anderson, Practical Process Research & Development—A Guide for    Organic Chemists, 2^(nd) ed., Academic Press, New York, 2012.-   Andreux et al., A method to identify and validate mitochondrial    modulators using mammalian cells and the worm C. elegans. Sci Rep;    4:5285; 2014.-   Andreux et al., Pharmacological approaches to restore mitochondrial    function. Nat Rev Drug Discov; 12(6):465-83; 2013.-   Artuso et al., Mitochondrial DNA metabolism in early development of    zebrafish (Danio rerio). Biochim Biophys Acta.; 1817(7):1002-11,    2012.-   Baraban et al., Pentylenetetrazole induced changes in zebrafish    behavior, neural activity and c-fos expression. Neuroscience. 2005;    131(3):759-68; 2005.-   Barton et al., Pharmacological characterization of the 6 Hz    psychomotor seizure model of partial epilepsy. Epilepsy Res;    47(3):217-27, 2001.-   Bialer and White, Key factors in the discovery and development of    new antiepileptic drugs. Nat Rev Drug Discov.; 9(1):68-82, 2010.-   Bindoff and Engelsen B A. Mitochondrial diseases and epilepsy.    Epilepsia.; 53:92-7, 2012.-   Bindoff and Engelsen, Mitochondrial cytopathies. In: Andermann F,    Guerrini R, Shorvon S D, editors. The Causes of Epilepsy: Common and    Uncommon Causes in Adults and Children. Cambridge: Cambridge    University Press; p. 147-57, 2011.-   Broughton et al., The complete sequence of the zebrafish (Danio    rerio) mitochondrial genome and evolutionary patterns in vertebrate    mitochondrial DNA. Genome Res.; 11(11):1958-67, 2001.-   Cheng et al., Retinoic acid protects against proteasome inhibition    associated cell death in SH-SY5Y cells via the AKT pathway.    Neurochem Int.; 62(1):31-42, 2013.-   Fei et al., Azaanthraquinone assembly from N-propargylamino quinone    via iodine-induced 6-endo-dig electrophilic cyclization. Org Biomol    Chem.; 8(18):4096-103, 2010.-   Fei et al., CuCl2-promoted 6-endo-dig chlorocyclization and    oxidative aromatization cascade: efficient construction of    1-azaanthraquinones from N-propargylaminoquinones. Org Lett.;    13(16):4208-11, 2011.-   Fieser and Fieser, The Reduction Potentials of Various    Naphthoquinones. J Am Chem Soc.; 57(3):491-4, 1935.-   Fieser, THE ALKYLATION OF HYDROXYNAPHTHOQUINONE I. ORTHO-ETHERS. J    Am Chem Soc.; 48(11):2922-37, 1926.-   Finsterer and Scorza, Effects of antiepileptic drugs on    mitochondrial functions, morphology, kinetics, biogenesis, and    survival. Epilepsy Res.; 136:5-11, 2017.-   Finsterer and Segall, Drugs interfering with mitochondrial    disorders. Drug Chem Toxicol.; 33(2):138-5, 2010.-   Fleming et al., Functional characterisation of the maturation of the    blood-brain barrier in larval zebrafish. PLoS One; 8(10):e77548,    2013.-   Franco et al., Challenges in the clinical development of new    antiepileptic drugs. Pharmacol Res.; 103:95-104, 2016.-   CN 101712648B. Synthesis method of azepine anthraquinone, 2010.-   Gornostaev et al., Synthesis of    13-alkylbenzo[f]isochromeno[4,3-b]indole-5,7,12(13H)-triones by    reaction of 2-alkylamino-1,4-naphthoquinones with ninhydrin. Russian    Journal of Organic Chemistry; 52(1):80-6, 2016.-   Ha and Park, Glutamate-induced oxidative stress, but not cell death,    is largely dependent upon extracellular calcium in mouse neuronal    HT22 cells. Neurosci Lett.; 393(2-3):165-9, 2006.-   Handbook of Pharmaceutical Salts: Properties, and Use, Stahl and    Wermuth Eds., Verlag Helvetica Chimica Acta, 2002.-   Hansen et al., Anticonvulsant and antiepileptogenic effects of GABAA    receptor ligands in pentylenetetrazole-kindled mice. Prog    Neuropsychopharmacol Biol Psychiatry; 28(1):105-13, 2004.-   Howe et al., The zebrafish reference genome sequence and its    relationship to the human genome. Nature; 496(7446):498-503, 2013.-   Hwang et al., 1998.-   Jeong et al., Functional and developmental analysis of the    blood-brain barrier in zebrafish. Brain Res Bull.; 75(5):619-28.    Epub 2008/03/22. doi: 10.1016/j.brainresbull.2007.10.043. PubMed    PMID: 18355638, 2008.-   Jiang and Wang, Gold(III)-Catalyzed 1,4-Nucleophilic Addition:    Facile Approach to Prepare 2-Amino-1,4-naphthalenedione and    6-Amino-5,8-quinolinedione Derivatives. Synlett.; 2009(07):1099-102,    2009.-   Jiang et al., Azaanthraquinone Assembly from N-Propargylamino    Quinone via a Au(I)-Catalyzed 6-endo-dig Cycloisomerization. J Org    Chem.; 75(12):4323-5, 2010.-   Josey et al., Structure-activity relationship study of vitamin k    derivatives yields highly potent neuroprotective agents. J Med Chem;    56(3):1007-22, 2013.-   Kimmel et al., Stages of embryonic development of the zebrafish. Dev    Dyn.; 203(3):253-310, 1995.-   Kumar et al., Synthesis of pharmacologically important    naphthoquinones and anticancer activity of 2-benzyllawsone through    DNA topoisomerase-II inhibition. Bioorg Med Chem.; 25(4):1364-73,    2017.-   Lewerenz et al., Activation of stimulatory heterotrimeric G proteins    increases glutathione and protects neuronal cells against oxidative    stress. J Neurochem.; 87(2):522-31, 2003.-   Lewerenz et al., Induction of Nrf2 and xCT are involved in the    action of the neuroprotective antibiotic ceftriaxone in vitro. J    Neurochem.; 111(2):332-43, 2009.-   Lheureux and Hantson, Carnitine in the treatment of valproic    acid-induced toxicity. Clin Toxicol (Phila).; 47(2):101-11, 2009.-   Lien et al., cSynthesis of 2-alkoxy 1,4-naphthoquinone derivatives    as antiplatelet, antiinflammatory, and antiallergic agents. Chem    Pharm Bull (Tokyo); 50(5):672-4, 2002.-   Loscher and Schmidt, Modern antiepileptic drug development has    failed to deliver: ways out of the current dilemma. Epilepsia.;    52(4):657-78, 2011.-   Loscher et al., New avenues for anti-epileptic drug discovery and    development. Nat Rev Drug Discov.; 12(10):757-76, 2013.-   Maher and Davis, The role of monoamine metabolism in oxidative    glutamate toxicity. J Neurosci.; 16(20):6394-401, 1996.-   Matagne and Klitgaard, Validation of corneally kindled mice: a    sensitive screening model for partial epilepsy in man. Epilepsy Res;    31(1):59-71, 1998.-   Mathiowitz et al., 1997.-   Matsumoto et al., Secondary elevation of extracellular    neurotransmitter amino acids in the reperfusion phase following    focal cerebral ischemia. J Cereb Blood Flow Metab.; 16(1):114-24,    1996.-   Metcalf et al., Development and pharmacologic characterization of    the rat 6 Hz model of partial seizures. Epilepsi; 58(6):1073-84,    2017.-   Milton et al., Rational design of quinones for high power density    biofuel cells. Chem Sci.; 6(8):4867-75, 2015.-   Mohanraj and Brodie, Outcomes in newly diagnosed    localization-related epilepsies. Seizure; 14(5):318-23, 2005.-   Morimoto and Koshland, Induction and expression of long- and    short-term neurosecretory potentiation in a neural cell line.    Neuron.; 5(6):875-80, 1990.-   Nadanaciva et al., Toxicity assessments of nonsteroidal    anti-inflammatory drugs in isolated mitochondria, rat hepatocytes,    and zebrafish show good concordance across chemical classes. Toxicol    Appl Pharmacol., 2013.-   Noebels et al., Jasper's Basic Mechanisms of the Epilepsies. 4th    edition ed. Bethesda (MD): National Center for Biotechnology    Information (US); 2012.-   Nogueira et al., “Syndromes associated with mitochondrial DNA    depletion.” Ital J Pediatr., 40:34, 2014.-   Novel tetracyclonaphthooxazole derivative and preparation method    thereof, 2015.-   Ogata et al., Unusual, chemoselective etherification of    2-hydroxy-1,4-naphthoquinone derivatives utilizing alkoxymethyl    chlorides: scope, mechanism and application to the synthesis of    biologically active natural product (±)-lantalucratin C.    Tetrahedron; 72(11):1423-32, 2016.-   Ohlow et al., Why Have Clinical Trials of Antioxidants to Prevent    Neurodegeneration Failed?—A Cellular Investigation of Novel    Phenothiazine-Type Antioxidants Reveals Competing Objectives for    Pharmaceutical Neuroprotection. Pharm Res.; 34(2):378-93, 2017.-   Peng et al., A 96-Well Screen Filter Plate for High-Throughput    Biological Sample Preparation and LC-MS/MS Analysis. Analytical    Chemistry; 78(1):343-8, 2006a.-   Peng et al., Fully Automated 96-Well Liquid-Liquid Extraction for    Analysis of Biological Samples by Liquid Chromatography with Tandem    Mass Spectrometry. Analytical Chemistry.; 73(3):708-14, 2001.-   Peng et al., Improved pharmacokinetic and bioavailability support of    drug discovery using serial blood sampling in mice. Journal of    pharmaceutical sciences; 98(5):1877-84, 2009.-   Peng et al., Particulate separation filters and methods. Google    Patents; 2006b.-   Perucca, Pharmacological and therapeutic properties of valproate: a    summary after 35 years of clinical experience. CNS Drugs.;    16(10):695-714, 2002.-   Petrova et al., Electrochemical properties of some naturally    occurring quinones. Journal of electroanalytical chemistry and    interfacial electrochemistry; 277(1-2):189-96, 1990.-   Poteet et al., Neuroprotective actions of methylene blue and its    derivatives. PLoS One; 7(10):e48279, 2012.-   Practical Process Research & Development, 2012.-   Rahn et al., Novel Vitamin K analogs suppress seizures in zebrafish    and mouse models of epilepsy. Neuroscience. 2014; 259C:142-54, 2013.-   Rahn et al., Novel Vitamin K analogs suppress seizures in zebrafish    and mouse models of epilepsy. Neuroscience; 259C:142-54, 2014.-   Reagan-Shaw et al., FASEB J., 22(3):659-661, 2008.-   Remington: The Science and Practice of Pharmacy, 21^(st) Ed.    Lippincott Williams and Wilkins, 2005.-   Remington's Pharmaceutical Sciences, 15th Edition, pages 1035-1038    and 1570-1580, 1975.-   Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company,    pp. 1289-1329, 1990.-   Rowley and White, Comparative anticonvulsant efficacy in the corneal    kindled mouse model of partial epilepsy: Correlation with other    seizure and epilepsy models. Epilepsy Res.; 92(2-3):163-9, 2010.-   Sagara and Schubert, The activation of metabotropic glutamate    receptors protects nerve cells from oxidative stress. J Neurosci.;    18(17):6662-71, 1998.-   Schriml et al., Human Disease Genes and Their Cloned Mouse    Orthologs: Exploration of the FANTOM2 cDNA Sequence Data Set. Genome    Research; 13(6b):1496-500, 2003.-   Schubert and Maher, An alternative approach to drug discovery for    Alzheimer's disease dementia. Future Med Chem.; 4(13):1681-8, 2012.-   Smith, March's Advanced Organic Chemistry: Reactions, Mechanisms,    and Structure, 7^(th) Ed., Wiley, 2013.-   Sreelatha et al., Synthesis and SAR study of novel anticancer and    antimicrobial naphthoquinone amide derivatives. Bioorg Med Chem    Lett.; 24(15):3647-51, 2014.-   Stables and Kupferberg, The NIH anticonvulsant drug development    (ADD) program: preclinical anticonvulsant. Molecular and cellular    targets for anti-epileptic drugs; 12:191, 1997.-   Stables et al., Therapy discovery for pharmacoresistant epilepsy and    for disease-modifying therapeutics: summary of the NIH/NINDS/AES    models II workshop. Epilepsia; 44(12):1472-8, 2003.-   Stewart et al., Polymerase gamma gene POLG determines the risk of    sodium valproate-induced liver toxicity. Hepatology.; 52(5):1791-6,    2010.-   Takenaga et al., 1998.-   Tan et al., Oxytosis: A novel form of programmed cell death. Curr    Top Med Chem.; 1(6):497-506, 2001.-   Tandon et al., Synthesis and evaluation of novel 1,4-naphthoquinone    derivatives as antiviral, antifungal and anticancer agents. Bioorg    Med Chem Lett; 14(11):2901-4, 2004.-   Tobaben et al., Bid-mediated mitochondrial damage is a key mechanism    in glutamate-induced oxidative stress and AIF-dependent cell death    in immortalized HT-22 hippocampal neurons. Cell Death Differ.;    18(2):282-92, 2011.-   Vafai et al., Natural Product Screening Reveals Naphthoquinone    Complex I Bypass Factors. PLoS One; 11(9):e0162686, 2016.-   Valente et al., The 1,4-naphthoquinone scaffold in the design of    cysteine protease inhibitors. Bioorg Med Chem; 15(15):5340-50, 2007.-   van Leyen et al., Novel lipoxygenase inhibitors as neuroprotective    reagents. J Neurosci Res.; 86(4):904-9, 2008.-   van Leyen et al., Proteasome inhibition protects HT22 neuronal cells    from oxidative glutamate toxicity. J Neurochem.; 92(4):824-30, 2005.-   Wang et al., Naphthoquinone-directed C—H annulation and C(sp(3))-H    bond cleavage: one-pot synthesis of tetracyclic naphthoxazoles. J    Org Chem.; 79(10):4553-60, 2014.-   Wang et al., Synthesis and Biological Evaluation of Lipophilic    1,4-Naphthoquinone Derivatives against Human Cancer Cell Lines.    Molecules; 20(7):11994-2015, 2015.-   Watanabe et al., In vivo assessment of the permeability of the    blood-brain barrier and blood-retinal barrier to fluorescent    indoline derivatives in zebrafish. BMC Neurosci.; 13(1):101, 2012.-   Wen et al., Alternative mitochondrial electron transfer as a novel    strategy for neuroprotection. J Biol Chem.; 286(18):16504-15, 2011.-   Westerfield, The zebrafish book. A guide for the laboratory use of    zebrafish (Danio rerio). 4th ed. Eugene: University of Oregon Press;    2000.-   Xie et al., A novel transgenic zebrafish model for blood-brain and    blood-retinal barrier development. BMC Dev Biol.; 10:76, 2010.-   Yang et al., The excitatory neurotransmitter glutamate stimulates    DNA repair to increase neuronal resiliency. Mech Ageing Dev.;    132(8-9):405-11, 2011.

1-54. (canceled)
 55. A compound of the formula:

or a pharmaceutically acceptable salt thereof.
 56. The compound of claim55 further defined as:


57. A pharmaceutical composition of claim 55 comprising: (A) a compoundof the formula:

or a pharmaceutically acceptable salt thereof; and (B) an excipient. 58.The pharmaceutical composition of claim 57, wherein the pharmaceuticalcomposition is formulated for oral, sublingual, intranasal, intravenous,subcutaneous, parenteral, inhalation, or aerosol delivery.
 59. Thepharmaceutical composition of claim 57, wherein the pharmaceuticalcomposition is formulated as a unit dose.